Toxicity, antifeedant and physiological effects of trans-anethole against Hyphantria cunea Drury (Lep: Arctiidae).

Plant secondary metabolites are currently known to interfere with basic metabolic, behavioral and physiological processes of insects. In the current study, the biological and physiological effects of trans-anethole were investigated against Hyphantria cunea Drury. The bioassay data demonstrated the high toxicity of trans-anethole against the fourth-instar larvae with the LC30, LC50 and LC90 values of 0.72, 1.41 and 7.20 µL/mL, respectively. Also the concentrations of LC30 and LC50 showed 53 and 87% feeding deterrency against the larvae. The biochemical experiments revealed that oral exposure of trans-anethole decreased the activities of digestive enzymes, acetylcholinesterase and the contents of energy reserves while, it induced the activities of detoxifying and antioxidant enzymes compared to control. In fact, trans-anethole induced the inhibition of digestion and AChE activities accompanied by imbalance in metabolic and oxidative processes so it may be recommended as a potent biopesticide in control of H. cunea populations.

Immunological interactions of Chilo suppressalis Walker (Lepidoptera: Crambidae) with the native entomopathogenic fungi.

Entomopathogenic fungi can attack many insect hosts and have been applied as the eco-friendly alternatives to synthetic chemicals for the control of pests. Insects have developed different defense systems encountering entomopathogens including humoral and cellular immune responses. In the present study, injection of some native entomopathogenic fungi to the Chilo suppressalis Walker larvae resulted in an enhancement of the cellular and antimicrobial defenses. The numbers of total and differential hemocytes increased rapidly in the first 3 and 6 h but those gradually reduced 12 and 24 h post-injections. The nodule formation and phenoloxidase activity increased at the time intervals after fungal infection. A similar trend was found in the transcription of antimicrobial peptides including attacin1 and 2, cecropin1 and 2, gallerimycin, defensin, lysozyme, and prophenoloxidase-activating proteinase-3 during infection fungi. In all cases, the target gene transcription was upper in the larvae injected by the fungi than that of control larvae. These results may elucidate better knowledge on the interaction of the fungi present in agroecosystems with the target insect pest.

Effect of thermal stress on the immune responses of Chilo suppressalis walker (Lepidoptera: Crambidae) to Beauveria bassiana.

Temperature is one of the important environmental elements affecting ecological fitness of insects through alterations in physiological systems. In the current study, a comparison was made on the cellular and humoral immune responses of the Chilo suppressalis larvae exposed to thermal stress (34 °C) and optimal rearing temperature (24 °C). Although total hemocyte count increased in the injected larvae by Beauveria bassiana, elevation of hemocyte numbers was significantly different in the larvae exposed to 34 °C for a short-time period compared to long-term exposure and control. A similar trend was observed in plasmatocyte and granulocyte counts as well as phenoloxidase activity. Gene expression of some antimicrobial peptides, including attacin1, attacin2, cecropin1, cecropin2, defensin, gallerimycin, lysozyme and prophenoloxidase-activating proteinase-3, was compared in the larvae exposed to thermal regimes and injection challenges. In all cases, expression of the target genes was relatively higher in the larvae injected by B. bassiana and short-term exposure at 34 °C. The present results confirmed that C. suppressalis could modulate the immune system in response to different thermal stress conditions mainly over a short period.

A Laboratory Study on the Modeling of Temperature-Dependent Development and Antioxidant System of Chilo suppressalis (Lepidoptera: Crambidae).

The developmental rates of Chilo suppressalis (Walker; Lepidoptera: Crambidae) were investigated at different constant temperatures of 11, 18, 22, 24, 26, 30, 32, 34, and 36°C to find out temperature thresholds, thermal requirements and larval antioxidant responses. The time to complete immature stages separately reduced by raising the rearing temperature except for eggs which showed no significant differences at 30-34°C. Data analysis by traditional and Ikemoto-Takai linear models determined Tmin of 10.0 and 9.92°C as well as thermal constants of 840.34 and 848.0 DD for the overall immature stages. The models including Analytis, Briere-2, Lactin-2, and Sharpe-Schoolfield-Ikemoto (SSI) calculated Tmin values of 9.15, 9.57, 10.0, and 11.37°C for overall immature stages while Tfast was found to be 33.8, 33.3, 33.7, and 33.0°C, respectively. Tmax was calculated as 35.12, 34.66, 34.56, 36.84, 34.11, and 35.15°C for Analytis, Briere-2, Lactin-2, SSI, Logan-6, and Logan-10. Topt using SSI was calculated as 24.42°C for total developmental time. The larvae exposure to 34°C in the short-term period demonstrated the highest activities of catalase, peroxidase, and superoxide dismutase compared to control (24°C). Activities of ascorbate peroxidase and glucose-6-phosphate hydrogenase also increased at 34°C in the short-term period for all preparations. The larvae exposed to 34°C in short-term period showed the highest amounts of Malondialdehyde and oxidized and reduced thiols (RSSR/RSH) ratio compared to control. These results may be useful to explain potentially ecological performance of C. suppressalis as the major pest of rice in Iran.

Rare parasitic copepods (Siphonostomatoida: Lernanthropidae) from Egyptian Red Sea fishes.

Two rare species of parasitic copepods belonging to the genus Lernanthropus de Blainville, 1822 (Siphonostomatoida: Lernanthropidae) are redescribed in detail, based on material collected from Red Sea fishes, caught at El-Tor, near Sharm El-Sheikh on the Red Sea coast of Egypt. Adult females of Lernanthropus sanguineus Song & Chen, 1976 were found on the gills of snapper Lutjanus fulviflamma (Forsskål). This species was known only from its original description based on material from Chinese waters. Adult females of Lernanthropus triangularis Pillai, 1963 were obtained from the gills of mojarra Gerres oyena (Forsskål). Both parasite species are new records for Egyptian Red Sea waters and both host records are new.

Parasitic copepods from Egyptian Red Sea fishes: Bomolochidae Claus, 1875.

Two species of parasitic copepods from the genus Bomolochus von Nordmann, 1832 (Cyclopoida: Bomolochidae) are redescribed in detail, based on material collected from the gills of Red Sea fishes. Host material was caught at El-tor, near Sharm El-Sheikh, and in the Gulf of Suez, Egypt. Both sexes of Bomolochus bellones Burmeister, 1835 were collected from the gills of a needlefish Tylosurus choram (Rüppell) caught in the Gulf of Suez. This is a new host record. The female is well characterised so only the male is described. Adult females of Bomolochus minus Lin & Ho, 2005 were obtained from the branchial cavities and gills of mojarra Gerres oyena (Forsskål). This species was known only from its original description in Taiwan, and this report constitutes a new host record and a significant range extension. Both parasite species are new records for Egyptian Red Sea waters.

Immune and metabolic responses of Chilo suppressalis Walker (Lepidoptera: Crambidae) larvae to an insect growth regulator, hexaflumuron.

Efficient control of Chilo suppressalis Walker is always controversial due to highly economic damage, resistance to insecticides and environmental pollutions. So, combination of safe pest controls e.g. biocontrol agents and insect growth regulators seems to be promising via integrated pest management program. Bioassay of hexaflumuron on 4th larval instars revealed concentrations of 44.34, 179.74 and 474.94µg/ml as LC10-50 values. Numbers of total hemocytes, plasmatocytes and granulocytes as well as phenoloxidase activity increased in the different time intervals following treatment by hexaflumuron. Combined effects of hexaflumuron and Beauveria bassiana Vuillemin also increased hemocyte numbers and phenoloxidase activity at different time intervals using all concentrations. Activities of general esterases assayed by α- and ß-naphtyl acetate and glutathione S-transferase using CDNB and DCNB increased 1-12h post-treatment. Activities of aspartate aminotransferase, alanine aminotransferase, γ-glutamyl transferase and aldolase increased in the larvae treated by hexaflumuron. However enhanced activity of lactate dehydrogenase was only obtained by treating 180 and 470µg/ml concentrations of hexaflumuron. Activities of ACP and ALP were found to be higher than control for all time intervals even 1-12h post-treatment. The amounts of HDL and LDL increased in the highest concentrations of hexaflumuron after 12-24h of post-treatment. Amount of triglyceride was higher than that of control after 1 and 3h but it was lower in other time intervals. Amounts of glycogen and protein were lower than those of control for all time intervals except for 6 and 12h of post-treatment in case of protein. Results of the current study revealed negative effects of hexaflumuron on intermediary metabolism of Chilo suppressalis but it increased the number of hemocytes and activity of phenoloxidase which are responsible for spore removal from hemolymph. It can be concluded that hexaflumuron is able to decrease survival and biological performance of C. suppressalis via intervening in intermediary metabolism but the given results showed incompatibility of the IGR with possible microbial control.

Purification and characterization of midgut α-amylase in a predatory bug, Andralus spinidens.

α-Amylases are widespread enzymes that catalyze endohydrolysis of long α-1,4-glucan chains such as starch and glycogen. The highest amylolytic activity was found in 5th instar nymphs and midgut of the predatory bug, Andrallus spinidens F. (Hemiptera: Pentatomidae). The α-amylase was purified following a three-step procedure. The purified α-amylase had a specific activity of 13.46 U/mg protein, recovery of 4.21, purification fold of 13.87, and molecular weight of 21.3 kDa. The enzyme had optimal pH and temperature of 7 and 45°C, respectively. Na+, Mn+, Mg2+, and Zn2+ significantly decreased activity of the purified α-amylase, but some concentrations of K+, Ca2+, and Cu2+ had the opposite effect. EDTA, EGTA, and DTC significantly decreased enzymatic activity, showing the presence of metal ions in the catalytic site of the enzyme. Kinetic parameters of the purified α-amylase showed a Km of 3.71% in starch and 4.96% for glycogen, suggesting that the enzyme had a higher affinity for starch.

A new parasitic copepod (Cyclopoida: Bomolochidae) from a ponyfish (Leiognathidae) caught in Egyptian Mediterranean waters, with a review of hosts and key to species of Nothobomolochus.

A new bomolochid copepod belonging to the genus Nothobomolochus Vervoort, 1962 is described from a Red Sea fish species, a ponyfish of the family Leiognathidae that has become established in the Eastern Mediterranean. The new species, N. leiognathicola n. sp., is based on material obtained from the gill chamber of the Red Sea immigrant ponyfish Leiognathus klunzingeri (Steindachner), caught in Egyptian waters off the Alexandria coast at Abuqir. A second new species, N. monodi n. sp., is established to accommodate some material previously described as N. denticulatus (Bassett-Smith, 1898), from the host Hemiramphus far Forsskål. A review of host records reveals that Nothobomolochus species utilise hosts representing five different orders, but are most commonly found on beloniform, clupeiform and perciform fishes. A newly constructed key to the 37 valid species of Nothobomolochus is presented.

Role of proteases in extra-oral digestion of a predatory bug, Andrallus spinidens.

Roles of salivary proteases in the extra-oral digestion of the predatory bug, Andrallus spinidens Fabricius (Hemiptera: Pentatomidae) were studied by using 2% azocasein as a general substrate and specific protease substrates, as well as synthetic and endogenous inhibitors. It was found that salivary glands of A. spinidens have two anterior, two lateral, and two posterior lobes. Azocasein was used to measure the activity of general proteases in the salivary glands using different buffer solutions. The enzyme had the highest activity at pH 8. General protease activity was highest at 40 °C and was stable for 6-16 hours. The use of specific substrates showed that trypsin-like, chymotrypsin-like, aminopeptidase, and carboxypeptidase are the active proteases present in salivary glands, by the maximum activity of trypsin-like protease in addition to their optimal pH between 8-9. Ca(2+) and Mg(2+) increased proteolytic activity about 216%, while other ions decreased it. Specific inhibitors including SBTI, PMSF, TLCK, and TPCK significantly decreased enzyme activity, as well as the specific inhibitors of methalloproteases including phenanthroline, EGTA, and TTHA. Extracted endogenous trypsin inhibitors extracted from potential prey, Chilo suppressalis, Naranga aenescens, Pieris brassicae, Hyphantria cunea, and Ephestia kuhniella, had different effects on trypsin-like protease activity of A. spinidens salivary glands. With the exception of C. suppressalis, the endogenous inhibitors significantly decreased enzyme activity in A. spinidens.

Diagnostic Utility of BAP1, EZH2 and Survivin in Differentiating Pleural Epithelioid Mesothelioma and Reactive Mesothelial Hyperplasia: Immunohistochemical Study.

Background: Epithelioid mesothelioma (EM) is the commonest subtype of malignant pleural mesothelioma. Its histopathological discrimination from reactive mesothelial hyperplasia (RMH) could be challenging. Thus, an immunohistochemical panel is mandatory for better discrimination. BAP1 is a newly identified diagnostic marker whose loss is specific to malignant mesothelioma. EZH2 overexpression is reported in different cancers, but its relation to BAP1 in malignant mesothelioma has not been fully understood. Survivin expression is said to be significantly higher in EM than in non-neoplastic pleural tissue, but its diagnostic utility as an immunohistochemical marker has not been thoroughly investigated in this field. To the best of our knowledge, no previous studies have been conducted to assess the diagnostic accuracy of the combined use of these three nuclear markers (BAP1, EZH2 and Survivin) in discriminating pleural EM from RMH. Methods: This retrospective study includes two groups: 81 cases of pleural EM and 67 cases of RMH, retrieved from the archives of Pathology Department of Ain Shams University Hospitals and Ain-Shams University Specialized Hospital during the period from January 2016 to December 2019. An immunohistochemical study was performed using BAP1, EZH2 and Survivin antibodies. Results: There were highly statistically significant relations between study groups as regards the studied markers (p = 0.001 for each). The specificity was 100% for all combinations of immunohistochemical markers. Sensitivity of any combination of the immunohistochemical markers used in this study was found to be higher than the sensitivity of any of these markers used individually. The combination of all three markers showed the highest diagnostic accuracy (95.9%) and the highest sensitivity (92.6%). However, the combination of Survivin and EZH2 yielded the same diagnostic accuracy and sensitivity. Conclusion: Adding EZH2, Survivin and BAP1 to the diagnostic IHC panel for differentiating pleural EM and RMH could enhance diagnostic sensitivity. Moreover, Survivin is a potentially promising marker in this context, especially when combined with EZH2.

Evaluation of the Prognostic Value of Solute Carrier Family 34 Member 2 "SLC34A2" in Papillary Thyroid Carcinoma: An Immunohistochemical Study.

BACKGROUND: Papillary thyroid carcinoma (PTC) usually has an indolent clinical course, yet a subset of patients might show an aggressive course. Thus, better stratification of at-risk patients is mandatory for proper management. Solute carrier family 34 member 2 (SLC34A2) is an independent prognostic indicator in several cancers. However, only a few studies have been conducted to evaluate the prognostic value of SLC34A2 in PTC, with none of them assessing its immunohistochemical (IHC) expression in a large cohort of patients with PTC or exploring its possible relationship with tumor progression. Aim of the Study. We aimed to evaluate the IHC expression of SLC34A2 in a large series of PTC patients, correlate its expression with established clinicopathological factors, and find any possible relationship between this marker and patient prognosis. Material and Methods. A total of 476 samples (including 238 samples of PTC and 238 samples of normal thyroid tissue) collected between 2002 and 2005 were extracted from the archives of the Pathology Lab, Ain Shams University Hospitals. IHC analysis was performed using an anti-SLC34A2 antibody. Follow-up data were obtained. RESULTS: High SLC34A2 expression significantly correlated with important adverse clinicopathological parameters of PTC-i.e., late tumor stage, positive extrathyroid extension, tumor size  >  4 cm, and age  ≥  55 years (p ≤ 0.001 for each). Kaplan-Meier analysis revealed that high SLC34A2 expression significantly correlated with shorter disease-free survival (DFS; p = 0.005), but not with overall survival (p = 0.111). Multivariate analysis showed SLC34A2 to be an independent prognostic factor affecting DFS. CONCLUSIONS: High SLC34A2 IHC expression correlated with adverse clinicopathological prognostic parameters. Furthermore, SLC34A2 was identified as an independent factor for DFS that could serve to improve risk stratification of PTC patients for better management.

Immunohistochemical expression of programmed death-ligand 1 and CD8 in glioblastomas.

BACKGROUND: Glioblastoma is the most aggressive primary malignant brain tumor in adults and is characterized by poor prognosis. Immune evasion occurs via programmed death-ligand 1 (PD-L1)/programmed death receptor 1 (PD-1) interaction. Some malignant tumors have responded to PD-L1/PD-1 blockade treatment strategies, and PD-L1 has been described as a potential predictive biomarker. This study discussed the expression of PD-L1 and CD8 in glioblastomas. METHODS: Thirty cases of glioblastoma were stained immunohistochemically for PD-L1 and CD8, where PD-L1 expression in glioblastoma tumor tissue above 1% is considered positive and CD-8 is expressed in tumor infiltrating lymphocytes. The expression of each marker was correlated with clinicopathologic parameters. Survival analysis was conducted to correlate progression-free survival (PFS) and overall survival (OS) with PD-L1 and CD8 expression. RESULTS: Diffuse/fibrillary PD-L1 was expressed in all cases (mean expression, 57.6%), whereas membranous PD-L1 was expressed in six of 30 cases. CD8-positive tumor-infiltrating lymphocytes (CD8+ TILs) had a median expression of 10%. PD-L1 and CD8 were positively correlated (p = .001). High PD-L1 expression was associated with worse PFS and OS (p = .026 and p = .001, respectively). Correlation of CD8+ TILs percentage with age, sex, tumor site, laterality, and outcomes were statistically insignificant. Multivariate analysis revealed that PD-L1 was the only independent factor that affected prognosis. CONCLUSIONS: PD-L1 expression in patients with glioblastoma is robust; higher PD-L1 expression is associated with lower CD8+ TIL expression and worse prognosis.

Effect of endoscopic variceal obliteration by band ligation on portal hypertensive gastro-duodenopathy: endoscopic and pathological study.

INTRODUCTION AND AIM: A few studies have shown that the degree of portal hypertensive gastropathy (PHG) and duodenopathy (PHD) has been worsening after the introduction of therapeutic endoscopic interventions. This study aimed to determine the impact of esophageal variceal eradication by endoscopic variceal ligation (EVL) on PHG and PHD using endoscopic and histopathologic assessment. METHODS: Fifty patients with esophageal varices for which EVL was indicated were included. EVL was carried out until complete variceal eradication was achieved. The degree of severity of PHG and PHD were recorded before and 4 weeks after variceal eradication. Biopsies were taken from various parts of the stomach and duodenum before and 4 weeks after variceal eradication. RESULTS: The whole Baveno score (4 vs. 2.5) increased significantly after variceal eradication when compared to those before eradication (p < 0.05). After obliteration, only 19 (38 %) patients had mild PHG versus 37 (74 %) before EVL, while severe PHG was found in 31 (62 %) patients versus 11 (22 %) before EVL and the difference was highly statistically significant. No significant changes were found regarding endoscopic PHD lesions before and after variceal eradication. Pathological changes as average blood vessel count, angiogenesis, ectasia and blood extravasation in stomach and duodenum significantly increased after EVL. Large esophageal varices (III-IV) and Baveno score (>1) at baseline endoscopy were independent risk factors for development of severe PHG after variceal obliteration (p < 0.05). CONCLUSION: PHG increased significantly, endoscopically and pathologically, after variceal obliteration by EVL. Although PHD did not significantly change as documented by endoscopy, pathological examination documented statistically significant changes in the duodenum after EVL.

Electrophoretic pattern and antibacterial activity of proteins from vicia faba seed extract.

Antibiotic resistance makes Antimierobial peptides (AMPs) agents an alternative for treatment of pathogenic diseases. They are isolated from various invertebrate animals, vertebrates and plants. The present study shows the electrophoretic pattern of protein and peptides from Vicia faba seed and reports our first attempt to study the antibacterial activity of Vicia faba seed extract. The crude extract electrophoresis was carried out on 12% SDS- PAGE gel. Antibacterial activity on E. Coli and B. Subtilis from hospital infection was tested and evaluated by measuring the inhibition zone diameter observation. The SDS- PAGE gel electrophoresis shows that the crude extract contains many proteins and peptides with different molecular weight. The inhibition zone was not observed in antibacterial properties tests. Thus, our experiments don't show any antibacterial activity on E. Coli and B. Subtilis from hospital isolates. Some other AMPs haven't also shown any antimicrobial properties on clinical trial. The antibacterial activity of the crude Vicia faba seed extract should also be tested on standard bacteria.

Dissociable components of rule-guided behavior depend on distinct medial and prefrontal regions.

Much of our behavior is guided by rules. Although human prefrontal cortex (PFC) and anterior cingulate cortex (ACC) are implicated in implementing rule-guided behavior, the crucial contributions made by different regions within these areas are not yet specified. In an attempt to bridge human neuropsychology and nonhuman primate neurophysiology, we report the effects of circumscribed lesions to macaque orbitofrontal cortex (OFC), principal sulcus (PS), superior dorsolateral PFC, ventrolateral PFC, or ACC sulcus, on separable cognitive components of a Wisconsin Card Sorting Test (WCST) analog. Only the PS lesions impaired maintenance of abstract rules in working memory; only the OFC lesions impaired rapid reward-based updating of representations of rule value; the ACC sulcus lesions impaired active reference to the value of recent choice-outcomes during rule-based decision-making.