AIMS: This article describes the qualitative and quantitative analyses of untypical compounds in the cuticular and internal lipids of four dipteran species. For isolated compounds, antimicrobial ...activity against 18 reference strains of bacteria and fungi was determined. METHODS AND RESULTS: In this study, gas chromatography (GC) combined with mass spectrometry (GC–MS) was used to analyse the surface and internal compounds of four fly species. Seven untypical compounds from both pre‐imaginal and imaginal stages of examined insects were identified. Azelaic acid (AA) was the most abundant, while phenylacetic and phenylpropionic acids occurred in lower concentration. Minor quantities of sebacic acid, 2‐methyl‐2‐hydroxybutanoic acid, tocopherol acetate and trace amounts of 2,4‐decadienal were also detected. Tocopherol acetate was found only in cuticular lipids of Musca domestica larvae. Each compound was tested against several species of fungi and bacteria by determining minimal inhibitory concentration (MIC). Human pathogenic fungi were also investigated. Phenylpropionic acid showed the greatest antifungal activity. Bacterial strains were insensitive to the presence of identified compounds, apart from 2,4‐decadienal which strongly inhibited bacterial growth. CONCLUSIONS: This is the first time that the chemical composition and the antimicrobial activity of untypical compounds in the cuticular and internal lipids of four fly species has been analysed. SIGNIFICANCE AND IMPACT OF THE STUDY: Determination of untypical compounds and their antimicrobial activity can effectively contribute to the knowledge concerning insect defence mechanisms.
Commonly used alkylimidazolium ionic liquids are poorly to negligibly biodegradable, and some are toxic, with the potential to poison typical biological test systems. Therefore, when ionic liquids ...are present in technological wastewaters they could break through classical wastewater treatment systems into natural waters and become potentially persistent pollutants. A recent study investigating different advanced oxidation processes found that the H
2O
2/UV system degraded dissolved imidazolium ionic liquids with the greatest efficiency. In the present study, high performance liquid chromatography was coupled with electrospray mass spectrometry to separate, analyse and identify degradation products following the treatment of ionic liquid solutions with H
2O
2 in the presence of UV irradiation. It was found that hydroxylation in short-chain entities occurred mainly within the ring moiety, whereas in the case of longer alkylated cations, oxidation of the alkyl chain yielded several products. The potential transformation products were identified structurally by MS/MS analysis and are discussed in the light of their putative toxicity and biodegradability.
Insect sulfakinins are pleiotropic neuropeptides with the homology to vertebrate gastrin/cholecystokinin peptide family. They have been identified in many insect species and affect different ...metabolic processes. They have a strong influence on feeding and digestion as well as on carbohydrate and lipid processing. Our study reveals that sulfakinins influence fatty acids composition in
Zophobas atratus
oenocytes and regulate insulin-like peptides (ILPs) level in these cells. Oenocytes are cells responsible for maintenance of the body homeostasis and have an important role in the regulation of intermediary metabolism, especially of lipids. To analyze the lipid composition in oenocytes after sulfakinins injections we used gas chromatography combined with mass spectrometry and for ILPs level determination an immunoenzymatic test was used. Because sulfakinin peptides and their receptors are the main components of sulfakinin signaling, we also analyzed the presence of sulfakinin receptor transcript (SKR2) in insect tissues. We have identified for the first time the sulfakinin receptor transcript (SKR2) in insect oenocytes and found its distribution more widespread in the peripheral tissues (gut, fat body and haemolymph) as well as in the nervous and neuro-endocrine systems (brain, ventral nerve cord,
corpora cardiaca/corpora allata
CC/CA) of
Z. atratus
larvae. The presence of sulfakinin receptor transcript (SKR2) in oenocytes suggests that observed effects on oenocytes lipid and ILPs content may result from direction action of these peptides on oenocytes.
Cardiorenal syndrome (CRS) is a group of pathophysiological disorders affecting heart and kidneys.
We present 44-year-old kidney transplant recipient with acute-on-chronic graft failure in the course ...of CRS due to acutely decompensated heart failure associated with severe aortic regurgitation successfully treated with aortic valve replacement. Because of graft failure progression and difficult to eradicate infections he was treated with dialysis and radical minimization of immunosuppression. After 74 days of renal replacement therapy the patient regained graft function after successful aortic valve replacement. The dialysis could be stopped and immunosuppressive therapy was reintroduced. Heart and renal function are stable and patient is doing well without dialysis for 3 years.
The return of kidney graft function can occur even after a long period of dialysis therapy due to improved cardiovascular function. Therefore, distinguishing an acute-on-chronic CRS subtype is mandatory to enable specific patient approach.
Gender influence on the clinical manifestations of Wilson’s Disease (WD) has been suggested; however, brain MRI pathology based on sexual dimorphism in WD has not yet been examined. The aim of this ...study was to analyse the effect of gender on brain MRI pathology according to the predominant form of WD. We retrospectively analysed the brain MR images of 204 newly diagnosed and untreated WD patients. The predominant form of the disease was neuropsychiatric (
n
= 105), hepatic (
n
= 67) or presymptomatic (
n
= 32). Overall, neuroimaging pathologies were found in 64.2 % WD patients. The clinical form analysis revealed significant gender-related differences. In the neuropsychiatric form, men presented with cerebellar atrophy and cortical brain atrophy more often than women (25/58 vs. 11/47;
p
< 0.05) and (23/58 vs. 12/47;
p
= 0.09), respectively. In contrast, women tended to present with globus pallidus lesions more often than men (25/47 vs. 20/58;
p
= 0.054). There were no gender differences observed in the hepatic form, but cortical brain atrophy presented more often in men than women (3/12 vs. 0/20;
p
< 0.05) in the presymptomatic form. According to our findings, there is a gender-dependent brain vulnerability to copper toxicity. We speculate that these differences are potentially related to an oestrogen protective effect and are due to differences in gender-related clinical forms.
.
Entomopathogenic fungi infect insects via penetration through the cuticle, which varies remarkably in chemical composition across species and life stages. Fungal infection involves the production ...of enzymes that hydrolyse cuticular proteins, chitin and lipids. Host specificity is associated with fungus–cuticle interactions related to substrate utilization and resistance to host‐specific inhibitors. The soil fungus Conidiobolus coronatus (Constantin) (Entomophthorales: Ancylistaceae) shows virulence against susceptible species. The larvae and pupae of Calliphora vicina (Robineau‐Desvoidy) (Diptera: Calliphoridae), Calliphora vomitoria (Linnaeus), Lucilia sericata (Meigen) (Diptera: Calliphoridae) and Musca domestica (Linnaeus) (Diptera: Muscidae) are resistant, but adults exposed to C. coronatus quickly perish. Fungus was cultivated for 3 weeks in a minimal medium. Cell‐free filtrate, for which activity of elastase, N‐acetylglucosaminidase, chitobiosidase and lipase was determined, was used for in vitro hydrolysis of the cuticle from larvae, puparia and adults. Amounts of amino acids, N‐glucosamine and fatty acids released were measured after 8 h of incubation. The effectiveness of fungal enzymes was correlated with concentrations of compounds detected in the cuticles of tested insects. Positive correlations suggest compounds used by the fungus as nutrients, whereas negative correlations may indicate compounds responsible for insect resistance. Adult deaths result from the ingestion of conidia or fungal excretions.
The composition of cuticular and internal n-alkanes in Lucilia sericata larvae, pupae, and male and female imagines were studied. The cuticular and internal lipid extracts were separated by ...HPLC-LLSD, after which the hydrocarbon fraction was identified by GC/MS in selected ion monitoring (SIM) and total ion current (TIC) modes. The cuticular lipids of the larvae contained seven n-alkanes from C23 to C31. The major n-alkane in L. sericata larvae was C29 (42.1%). The total cuticular n-alkane content in the cuticular lipids was 31.46 μg g−1 of the insect body. The internal lipids of L. sericata larvae contained five n-alkanes ranged from C25 to C31. The most abundant compound was C27 (61.71 μg g−1 of the insect body). Eighteen n-alkanes from C14 to C31 were identified in the cuticular lipids of the pupae. The most abundant n-alkanes ranged from C25 to C31; those with odd-numbered carbon chains were particularly abundant, the major one being C29:0 (59.5%). Traces of eight cuticular n-alkanes were present. The internal lipids of L. sericata pupae contained five n-alkanes, ranging from C25 to C31. The cuticular lipids of female imagines contained 17 n-alkanes from C12 to C30. Among the cuticular n-alkanes of females, C27 (47.5%) was the most abundant compound. Four n-alkanes, with only odd-numbered carbon chains, were identified in the internal lipids of females. The lipids from both sexes of L. sericata had similar n-alkane profiles. The cuticular lipids of adult males contained 16 n-alkanes ranging from C13 to C31. C27 (47.9%) was the most abundant cuticular n-alkanes in males. The same n-alkanes only with odd-numbered carbon chains and in smaller quantities of C27 (0.1%) were also identified in the internal lipids of males. The highest amounts of total cuticular n-alkanes were detected in males and females of L. sericata (330.4 and 158.93 μg g−1 of the insect body, respectively). The quantities of total cuticular alcohols in larvae and pupae were smaller (31.46 μg g−1 and 42.08 μg g−1, respectively). The internal n-alkane contents of larvae, pupae, and male and female imagines were significantly higher than the cuticular n-alkane contents (153.53, 99.60, 360.06 and 838.76 μg g−1 of the insect body, respectively).
Epicuticular lipids in many terrestrial arthropods consist of vast numbers of polar and non-polar aliphatic compounds, which are mainly responsible for the water balance in these animals but can also ...affect conidia germination of entomopathogenic fungi. In this work the qualitative and quantitative profiles of cuticular fatty acids from three insect species differing in their susceptibility to fungal infection were studied. In an innovative approach, laser light scattering detection was coupled with HPLC in order to identify the non-chromophoric chemicals usually present in cuticular extracts. The acids identified contained from 5 to 20 carbon atoms in the alkyl chain and included unsaturated entities such as C
16:1, C
18:1, C
18:2, C
18:3 and C
20:1. There was a marked dominance of acids containing 16–18 carbon atoms. The relative contents of fatty acids in the extracted waxes varied from trace amounts to 44%. Cuticular fatty acids profile of
Calliphora vicina (species resistant to fungal infection) significantly differs from profiles of
Dendrolimus pini and
Galleria mellonella (both species highly susceptible to fungal infection). The major difference is the presence of C
14:0, C
16:1 and C
20:0 in the cuticle of
C. vicina. These three fatty acids are absent in the cuticle of
D. pini while
G. mellonella cuticle contains their traces. The concentrations of four fatty acids dominating in the
G. mellonella larval cuticle (C
16:0, C
18:0, C
18:1 and C
18:2) were found to fluctuate during the final larval instar and correlate with fluctuations in the susceptibility of larvae to fungal infection. The possible role of cuticular fatty acids in preventing fungal infection is discussed.
Transcription factor IIH (TFIIH) is essential for both transcription and nucleotide excision repair (NER). DNA lesions are initially detected by NER factors XPC and XPE or stalled RNA polymerases, ...but only bulky lesions are preferentially repaired by NER. To elucidate substrate specificity in NER, we have prepared homogeneous human ten-subunit TFIIH and its seven-subunit core (Core7) without the CAK module and show that bulky lesions in DNA inhibit the ATPase and helicase activities of both XPB and XPD in Core7 to promote NER, whereas non-genuine NER substrates have no such effect. Moreover, the NER factor XPA activates unwinding of normal DNA by Core7, but inhibits the Core7 helicase activity in the presence of bulky lesions. Finally, the CAK module inhibits DNA binding by TFIIH and thereby enhances XPC-dependent specific recruitment of TFIIH. Our results support a tripartite lesion verification mechanism involving XPC, TFIIH, and XPA for efficient NER.
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•Human ten-subunit TFIIH and its seven-subunit Core7 were purified to homogeneity•Bulky lesions in DNA inhibit both the XPB and XPD helicase activities of Core7•XPA further inhibits the Core7 helicase activity in the presence of a bulky lesion•The CAK module enhances lesion-dependent recruitment of TFIIH by XPC in NER
Nucleotide-excision repair (NER) is essential for mutation avoidance and genome maintenance. However, NER also contributes to drug resistance and reduced efficacy of chemotherapy. Elucidating the molecular mechanism of DNA lesion recognition in NER leads to better understanding of genetic disorders and improved diagnosis and treatment of a number of diseases.
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