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The present study aimed to perform molecular identification of lepidopteran larvae from canopy fogging including gut-content analyses. A total of 130 lepidopteran larvae were selected from 37 fogging samples at the Panguana station, district Yuyapichis, province Puerto Inca, department Huánuco, Peru. Target trees were pre-identified and subsequently submitted to molecular confirmation of identity with three markers (rbcL, psbA and trnL-F). The COI gene of 119 lepidopteran larvae was successfully sequenced and found to belong to 92 species: Comparison of DNA barcodes with the reference database of adult moths resulted in 65 (55%) matches at species level, 32 (27%) at genus level, 19 (16%) at subfamily or family level, three just to order level. Three larvae could not be assigned to a family. For these larvae the fogged target tree now suggests a potential host-plant relationship. Molecular gut content analysis, based on High-Throughput-Sequencing was successfully tested for ten larvae corroborating feeding on the target plant in some cases but elucidating several other cases of potential 'alternative feeding'. We propose a larger-scale approach using this rapid and efficient method including molecular gut-content analyses for comprehensively testing the ratio of 'alternative feeders' and pitfalls caused by collateral fogging of larvae from neighboring trees. [ABSTRACT FROM AUTHOR]

Background: Here, we present applications and workflows of DNA metabarcoding utilizing next-generation sequencing (NGS) for identification of animal species present in environmental samples. All projects took place within the framework [...]

Background: Biodiversity patterns are inherently complex and difficult to comprehensively assess. Yet, deciphering shifts in species composition through time and space are crucial for successful management of ecosystem services, as [...]

Aim: To determine the effects of vitamin E on transforming growth factor-beta2 (TGF-β2)-induced cellular changes in cultured human trabecular meshwork (TM) cells. Materials and Methods : Human TM cells were pre-treated with different concentrations of vitamin E. Afterwards, cells were exposed to 1.0 ng/ml TGF-β2 for 24 h. Expressions of the heat shock protein αB-crystallin, the extracellular matrix (ECM) component fibronectin and the ECM-degrading enzyme matrix metalloproteinase-2 (MMP-2) were examined by real-time polymerase chain reaction (PCR) analyses. The cytoskeleton was investigated by phalloidin staining. Results : TGF-β2 increased the expressions of αB-crystallin and fibronectin and reduced the levels of MMP-2. TGF-β2 induced the formation of actin stress fibers and cross-linked actin networks. Pre-treatment with different concentrations of vitamin E reversed the TGF-β2-induced cellular changes in cultured human TM cells. Conclusions : TGF-β2-mediated changes in human TM cells could be reduced by pre-treatment with vitamin E. Therefore, it may be speculated that increasing the antioxidative capacity may help to lower the incidence of characteristic glaucomatous changes in the TM. [ABSTRACT FROM AUTHOR]

Background/Aims: The goal of the present study was to determine whether transforming growth factor-β2 (TGF-β2)- and oxidative stress-induced cellular changes in cultured human optic nerve head (ONH) astrocytes could be reduced by pretreatment with the antioxidant α-lipoic acid (LA). Methods: Cultured ONH astrocytes were treated with 1.0 ng/ml TGF-β2 for 24 h or 200 μM hydrogen peroxide (H2O2) for 1 h. Lipid peroxidation was measured by a decrease in cis-parinaric acid fluorescence. Additionally, cells were pretreated with different concentrations of LA before TGF-β2 or H2O2 exposure. Expressions of the heat shock protein (Hsp) αB-crystallin and Hsp27, the extracellular matrix (ECM) component fibronectin and the ECM-modulating protein connective tissue growth factor (CTGF) were examined with immunohistochemistry and real-time PCR analysis. Results: Both TGF-β2 and H2O2 increased lipid peroxidation. Treatment of astrocytes with TGF-β2 and H2O2 upregulated the expression of αB-crystallin, Hsp27, fibronectin and CTGF. Pretreatment with different concentrations of LA reduced the TGF-β2- and H2O2-stimulated gene expressions. Conclusion: We showed that TGF-β2- and H2O2-stimulated gene expressions could be prevented by pretreatment with the antioxidant LA in cultured human ONH astrocytes. Therefore, it is tempting to speculate that the use of antioxidants could have protective effects in glaucomatous optic neuropathy. Copyright © 2012 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

The effects of novel perennial energy crops on arthropod abundance and diversity were tested using three field trials in Bavaria. Established cup plant (Silphium perfoliatum), Virginia mallow (Sida hermaphrodita) and tall wheat grass (Thinopyrum ponticum) plots were sampled in autumn using a modified leaf blower and their arthropod faunas compared with that recorded in maize plots. At the time of sampling, maize, cup plant and Virginia mallow had already been harvested and samples from the remaining stubble were compared with those from open ground. Collembola were the most abundant group. The highest abundance and diversity of arthropods was recorded in tall wheat grass, with Hemiptera and Aranea especially benefitting from the green and taller vegetation. Post-harvest cup plant and Virginia mallow plots harboured a more diverse and more abundant arthropod assemblage than maize plots. Stubble was identified as a suitable habitat for arthropods in agricultural fields. In conclusion, the integration of perennial energy crops into existing biogas production systems could play an important role in supporting arthropods in agricultural landscapes by providing crucial structures and resources, such as the grassy vegetation of tall wheat grass in autumn and the long-lasting stubbles of cup plant and Virginia mallow. [ABSTRACT FROM AUTHOR]

Background: Dragonflies and damselflies (Odonata) are important components in biomonitoring due to their amphibiotic lifecycle and specific habitat requirements. They are charismatic and popular insects, but can be challenging to identify despite large size and often distinct coloration, especially the immature stages. DNA-based assessment tools rely on validated DNA barcode reference libraries evaluated in a supraregional context to minimize taxonomic incongruence and identification mismatches., Methods: This study reports on findings from the analysis of the most comprehensive DNA barcode dataset for Central European Odonata to date, with 103 out of 145 recorded European species included and publicly deposited in the Barcode of Life Data System (BOLD). The complete dataset includes 697 specimens (548 adults, 108 larvae) from 274 localities in 16 countries with a geographic emphasis on Central Europe. We used BOLD to generate sequence divergence metrics and to examine the taxonomic composition of the DNA barcode clusters within the dataset and in comparison with all data on BOLD., Results: Over 88% of the species included can be readily identified using their DNA barcodes and the reference dataset provided. Considering the complete European dataset, unambiguous identification is hampered in 12 species due to weak mitochondrial differentiation and partial haplotype sharing. However, considering the known species distributions only two groups of five species possibly co-occur, leading to an unambiguous identification of more than 95% of the analysed Odonata via DNA barcoding in real applications. The cases of small interspecific genetic distances and the observed deep intraspecific variation in Cordulia aenea (Linnaeus, 1758) are discussed in detail and the corresponding taxa in the public reference database are highlighted. They should be considered in future applications of DNA barcoding and metabarcoding and represent interesting evolutionary biological questions, which call for in depth analyses of the involved taxa throughout their distribution ranges., Competing Interests: Jérôme Morinière is founder & CEO of AIM Advanced Identification Methods GmbH. All authors declare that they have no competing interests., (© 2021 Geiger et al.)

Background: Biodiversity patterns are inherently complex and difficult to comprehensively assess. Yet, deciphering shifts in species composition through time and space are crucial for efficient and successful management of ecosystem services, as well as for predicting change. To better understand species diversity patterns, Germany participated in the Global Malaise Trap Program, a world-wide collection program for arthropods using this sampling method followed by their DNA barcode analysis. Traps were deployed at two localities: "Nationalpark Bayerischer Wald" in Bavaria, the largest terrestrial Natura 2000 area in Germany, and the nature conservation area Landskrone, an EU habitats directive site in the Rhine Valley. Arthropods were collected from May to September to track shifts in the taxonomic composition and temporal succession at these locations., New Information: In total, 37,274 specimens were sorted and DNA barcoded, resulting in 5,301 different genetic clusters (BINs, Barcode Index Numbers, proxy for species) with just 7.6% of their BINs shared. Accumulation curves for the BIN count versus the number of specimens analyzed suggest that about 63% of the potential diversity at these sites was recovered with this single season of sampling. Diversity at both sites rose from May (496 & 565 BINs) to July (1,236 & 1,522 BINs) before decreasing in September (572 & 504 BINs). Unambiguous species names were assigned to 35% of the BINs (1,868) which represented 12,640 specimens. Another 7% of the BINs (386) with 1,988 specimens were assigned to genus, while 26% (1,390) with 12,092 specimens were only placed to a family. These results illustrate how a comprehensive DNA barcode reference library can identify unknown specimens, but also reveal how this potential is constrained by gaps in the quantity and quality of records in BOLD, especially for Hymenoptera and Diptera. As voucher specimens are available for morphological study, we invite taxonomic experts to assist in the identification of unnamed BINs.

Background/aims: The goal of the present study was to determine whether transforming growth factor-β(2) (TGF-β(2))- and oxidative stress-induced cellular changes in cultured human optic nerve head (ONH) astrocytes could be reduced by pretreatment with the antioxidant α-lipoic acid (LA)., Methods: Cultured ONH astrocytes were treated with 1.0 ng/ml TGF-β(2) for 24 h or 200 µM hydrogen peroxide (H(2)O(2)) for 1 h. Lipid peroxidation was measured by a decrease in cis-parinaric acid fluorescence. Additionally, cells were pretreated with different concentrations of LA before TGF-β(2) or H(2)O(2) exposure. Expressions of the heat shock protein (Hsp) αB-crystallin and Hsp27, the extracellular matrix (ECM) component fibronectin and the ECM-modulating protein connective tissue growth factor (CTGF) were examined with immunohistochemistry and real-time PCR analysis., Results: Both TGF-β(2) and H(2)O(2) increased lipid peroxidation. Treatment of astrocytes with TGF-β(2) and H(2)O(2) upregulated the expression of αB-crystallin, Hsp27, fibronectin and CTGF. Pretreatment with different concentrations of LA reduced the TGF-β(2)- and H(2)O(2)-stimulated gene expressions., Conclusion: We showed that TGF-β(2)- and H(2)O(2)-stimulated gene expressions could be prevented by pretreatment with the antioxidant LA in cultured human ONH astrocytes. Therefore, it is tempting to speculate that the use of antioxidants could have protective effects in glaucomatous optic neuropathy., (Copyright © 2012 S. Karger AG, Basel.)

Purpose: Recent studies have revealed an accumulation of senescent cells in the outflow pathways in primary open-angle glaucoma (POAG). Transforming growth factor (TGF)-β2 is thought to be involved in the pathologic changes of the trabecular meshwork (TM) of POAG eyes. The goal of this study was to determine whether TGF-β2 triggers senescence-associated changes in human TM cells in vitro., Methods: Cultured human TM cells were exposed to 1.0 ng/mL TGF-β2 for 12, 24, and 48 hours. Senescence-associated β-galactosidase (SA-β-Gal) activity was investigated by histochemical staining. Lipid peroxidation was assessed after TGF-β2 treatment. Levels of apolipoprotein J (Apo J), SM22, and osteonectin (SPARC) mRNA were determined by real-time PCR analysis. Furthermore, the effects of antioxidants on these TGF-β2-mediated changes were tested. Induction of senescence-related signal transduction proteins (p16, p21, and pRb) was examined by real-time PCR and Western blot analysis., Results: TGF-β2 increased SA-β-Gal activity, lipid peroxidation, and the mRNA expressions of Apo J, SM22, and SPARC. These TGF-β2-induced changes were attenuated by antioxidants. TGF-β2 increased p16 mRNA and protein expression, which was paralleled by a downregulation of pRb protein. There was no effect on p21 mRNA and protein expression after exposure to TGF-β2., Conclusions: TGF-β2 induces senescence-associated TM changes and activates the senescence-related p16-pRb signal transduction pathway in vitro. Thus, minimizing TGF-β2 levels may help to prevent the ageing process in the TM as seen in POAG.

Purpose: Histologic studies have previously demonstrated increased expression of small heat shock proteins (Hsps) in reactive optic nerve head (ONH) astrocytes of patients with glaucoma. Transforming growth factor (TGF)-beta2 and hydrogen peroxide (H(2)O(2)) are known to induce ONH astrocyte reactivation. The goal of the present study was to determine whether new potentially involved Hsps, such as Hsp32, -47, -60, and -70, are expressed in the reactivation process of ONH astrocytes mediated by TGF-beta2 and H(2)O(2)., Methods: Cultured human ONH astrocytes were treated with 1.0 ng/mL TGF-beta2 for up to 48 hours. In addition, the cells were exposed to 100, 200, or 400 microM H(2)O(2) for 1 hour. Expression of Hsp32, -47, -60, and -70 was examined by immunohistochemistry, real-time PCR, and Western blot analyses., Results: Treatment with TGF-beta2 increased Hsp32 after 4 and 6 hours, whereas Hsp47 was upregulated after treatment with TGF-beta2 for 12, 24, and 48 hours. Exposure of the cells to H(2)O(2) could increase both Hsp32 and -47. No significant effects on the expression of Hsp60 and -70 were observed after treatment of the cells with TGF-beta2 or H(2)O(2)., Conclusions: TGF-beta2 increased Hsp32 after short-term treatment and Hsp47 after longer periods in cultured human ONH astrocytes. H(2)O(2) increased both Hsp32 and -47 levels. No effects on Hsp60 and -70 levels were induced by TGF-beta2 and H(2)O(2). These results may provide further insights into the cellular stress responses of reactive human ONH astrocytes. Further extensive studies are needed to examine the potential roles of Hsps in the ONH of glaucomatous eyes.