Anti-Diabetic Effects and Molecular Mechanisms of Amide Alkaloids from Piper longum Based on Network Pharmacology Integrated with Cellular Assays

Piper longum is a well-known spice and traditional medicine. It was revealed to possess anti-diabetic activity, but few information about its active component and underlying mechanism could be available. In this study, retrofractamides A ( 1 ) and C ( 2 ) isolated from P. longum showed potent inhibitory activity against PTP1B. Therefore, the potential mechanism was predicted by network pharmacology and molecular docking. PI3K/AKT was obtained as the most remarkable pathway against type 2 diabetes mellitus (T2DM), and AKT1 and GSK3β were yielded as the top two core targets of retrofractamides A ( 1 ) and C ( 2 ). Molecular docking of compounds with AKT1 and GSK3β showed strong binding affinity between them. Additionally, cellular experiments with a L6 cell model was conducted to further verify the above predictions. Results indicated that retrofractamides A ( 1 ) and C ( 2 ) exerted anti-diabetic effect via activating PI3K/AKT pathway, and they promoted glucose consumption, glucose uptake, glycogen synthesis and glycolysis.     

The kinetics of the active and de-energized transport of O-methyl glucose in Ustilago maydis

The kinetics of the uptake and efflux of 3-O-methyl-glucose in sporidia of Ustilago maydis were measured, both in active cells and in cells whose metabolic activity had been inhibited by azide and iodoacetate.The de-energized transport system proved to be carrier mediated with apparent affinity constants 13 ± 2 mM outside (K₀) and 18 ± 2 mM inside (K₁). The apparent maximum rate constants for the same system were 0.66 ± 0.05 mmol/l cell water per min for uptake (V₊ and 0.53 ± 0.04 mmol/l cell water per min for efflux (V_-. For the active system K₀ = 0.08 ± 0.01, K₁ > 40, V₊ = 9.7 ± 0.5 and V₋ 1.1 ± 0.9 (in equivalent units). These results are discussed in the context of the carrier mechanism as proposed by Regen and Morgan (Regen, D.M. and Morgan, H.E. (1964) Biochim. Biophys. Acta 79, 151–166).The antifungal compound carboxin had no effect on de-energized transport but was shown to decrease both K₀ and V₊ in the active system. Phloretin and phlorizin were also found to be without effect on de-energized cells but the former enhanced while the latter inhibited active uptake.     

Inferring species boundaries using acoustic and morphological data in the ground cricket genus Gymnogryllus (Orthoptera: Grylloidea: Gryllinae)

An important function of song production by male crickets is to attract conspecific females. These sound signals can be used to infer species boundaries as they can provide indirect evidence for reproductive isolation. However, many studies of orthopteran diversity in South-east Asia are based mainly on morphology and only occasionally acoustics. As such, there is a lack of information on how acoustic data can be congruent with morphological data when used to delineate species. Crickets of the genus Gymnogryllus (Grylloidea, Gryllidae), are such an example. Gymnogryllus are relatively speciose, but their calling songs have not been studied. We collected specimens and calling songs of five Gymnogryllus species from South-east Asia. The acoustic parameters of the calls, along with male tegminal venation and morphology genitalia, were compared. All data types showed congruency in distinguishing G. sylvestris and G. leucostictus from each other and from the other species. Inferring species boundaries for G. angustus, G. malayanus, and G. unexpectus using acoustics and tegminal morphometry proves to be more challenging. While acoustics, tegminal morphometry, and genital morphology are likely to be useful for inferring species of Gymnogryllus from different species groups, greater coverage of taxa is needed to resolve taxonomy of closely related Gymnogryllus.     

Point Mutation Approach to Reduce Antigenicity of Interferon Beta

Interferon beta (IFNβ) is naturally occurring cytokine made and secreted by immune cells in response to stimuli. Non-glycosylated interferon beta Ser     

Reproductive biology of the Mugger Crocodile,Crocodylus palustris,in Iran (Reptilia: Crocodylidae)

A small population of Mugger crocodiles occurs in the southeastern part of Iran, in Sistan & Baluchestan Province, at the westernmost global range of the species. The peak nesting season is May and consequently the eggs hatch in July, but the nesting season can be extended. In four nests found in the wild, 24, 26, 21 and 13 eggs were counted. A 2.2 m long female, which fell in an overflow pond, laid 21 eggs on a concrete surface exposed to the air. A female kept in captivity laid 18 and 25 eggs in two consecutive years. The mean size for the eggs in the last three nests was calculated as 75.7 x 47.7 mm and 91.27 g in weight. The mean total length and weight for 19 hatchlings was 30.47 cm (29.2–33.9 cm) and 84.3 g (66.2 to 90.3 g) respectively. Based on the number of observed hatchlings compared with the clutch size, it seems that hatchlings have a high mortality rate in the early stages after hatching as a result of natural threats.     

Importance of calcium-binding site 2 in simian virus 40 infection

The exposure of molecular signals for simian virus 40 (SV40) cell entry and nuclear entry has been postulated to involve calcium coordination at two sites on the capsid made of Vp1. The role of calcium-binding site 2 in SV40 infection was examined by analyzing four single mutants of site 2, the Glu160Lys, Glu160Arg, Glu157Lys (E157K), and Glu157Arg mutants, and an E157K-E330K combination mutant. The last three mutants were nonviable. All mutants replicated viral DNA normally, and all except the last two produced particles containing all three capsid proteins and viral DNA. The defect of the site 1-site 2 E157K-E330K double mutant implies that at least one of the sites is required for particle assembly in vivo. The nonviable E157K particles, about 10% larger in diameter than the wild type, were able to enter cells but did not lead to T-antigen expression. Cell-internalized E157K DNA effectively coimmunoprecipitated with anti-Vp1 antibody, but little of the DNA did so with anti-Vp3 antibody, and none was detected in anti-importin immunoprecipitate. Yet, a substantial amount of Vp3 was present in anti-Vp1 immune complexes, suggesting that internalized E157K particles are ineffective at exposing Vp3. Our data show that E157K mutant infection is blocked at a stage prior to the interaction of the Vp3 nuclear localization signal with importins, consistent with a role for calcium-binding site 2 in postentry steps leading to the nuclear import of the infecting SV40.     

Cytomorphological and immunocytochemical study of non-Hodgkin's lymphoma in pleural effusion and ascitic fluid

INTRODUCTION: Non-Hodgkin's lymphoma (NHL) is often complicated by pleural effusion and ascites. The present study is an attempt to categorize the lymphomatous effusions according to the WHO classification, using archival material. METHODS: May-Grünwald-Giemsa and Papanicolaou-stained smears of 31 lymphomatous effusion specimens were reviewed. Of these, detailed cytological assessment was done on 12 pleural effusions and ten ascitic fluid specimens from 22 patients using the WHO lymphoma classification system. Immunocytochemical studies were performed in 21 specimens. RESULTS: Based on cytomorphological features, the 22 lymphomatous effusion specimens were categorized into lymphoplasmacytoid lymphoma (1), follicle centre cell (FCC) grade-1 (centrocytic) lymphoma (3), FCC grade-2 (centrocytic-centroblastic) lymphoma (3), FCC grade-3 (centroblastic) lymphoma (4), large cell immunoblastic lymphoma (4), lymphoblastic lymphoma (2), anaplastic large cell lymphoma (3) and miscellaneous types (2). Immunocytochemically, the lymphoma cells were T-cell (positive for CD3) and B-cell type (CD20 positive) in five and six cases respectively. CONCLUSION: Cytological examination of pleural effusion and ascitic fluid samples, supported by immunocytochemical studies, may be useful for the classification of lymphomas under the WHO system.