Background
Most recently described human platelet antigens (
HPAs
) have been detected in cases of fetomaternal alloimmune thrombocytopenia (
FMAIT
) where the mother has been immunized against a ...low‐frequency antigen that the fetus has inherited from the father. Low‐frequency antigens are not represented in normal panel platelets (
PLTs
) and antibody detection and identification in such cases requires incubation of maternal serum with paternal
PLTs
and definition of the causative mutation.
Study Design and Methods
A suspected case of
FMAIT
was investigated for
PLT
‐specific antibodies using a panel of both
HPA
‐typed and paternal
PLTs
.
HPA
typing was performed by polymerase chain reaction with sequence‐specific primers and further
DNA
analysis was performed using direct sequencing of the coding regions of the
ITGA
2
B
and
ITGB
3
genes.
Results
Maternal antibodies reactive only with paternal
PLTs
were localized to glycoprotein (
GP
)
IIb
/
IIIa
using the monoclonal antibody immobilization of
PLT
antibody assay. A single‐nucleotide polymorphism was detected in
E
xon 23 of
ITGA
2
B
in the father and affected child, which predicted a
V
740
L
substitution in the mature protein. Recombinant
V
740
L
mutated
GPIIb
expressed in
HEK
293 cells was specifically recognized by maternal antibodies. The polymorphism was not detected either in the mother or in a cohort of 100 donors.
Conclusion
The
V
740
L
polymorphism defines a new low‐frequency antigen implicated in two cases of
FMAIT
in a single family. Low‐frequency
HPAs
are clinically important and their elucidation requires both crossmatch studies and gene sequencing in cases where there is strong clinical evidence of
FMAIT
but initial laboratory investigations do not support the diagnosis.
IntroductionAKI is very common in paediatric cardiac surgery patients. The diagnosis relies on changes in serum creatinine and urine output, which lack specificity and sensitivity especially in ...newborns and premature infants. Diagnostic changes also occur late in the injury phase when interventions have limited effectiveness. Microvesicles (MV) are small particles (<1μm) in plasma that often contain miRNA. Their levels have been shown to be altered in animal models of AKI where they regulate injury responses.HypothesisMV and associated miRNA provide specific and early-detection markers for AKI and a tool to investigate AKI pathogenesis.MethodsStudy subjects were 24 children (<17 yrs), undergoing cardiac surgery with cardiopulmonary bypass (CPB) without pre-existing inflammatory state, AKI or extracorporeal life support. AKI was diagnosed following the KDIGO definition. Blood and urine samples were collected pre-operatively and at 6 and 24 hours. Cytokines were measured on a MAGPIX platform. Leukocyte and platelets activation, and MV derivation was assessed by flow cytometry and NanoSight device. miRNAs were isolated from MV preps and analyzed by RT-PCR using Applied Biosystems kits.ResultsAKI was diagnosed in 14 patients and was confirmed by urinary NGAL increase at 6 hours after the operation (25±3.45 ng/ml vs 10.44±2.9 ng/ml; p 0.006). Patients with AKI were younger, underwent longer CPB, received higher volumes of red blood cells, and required higher inotrope support (Table). AKI subjects had increased levels of monocyte and B1-integrin MV. That correlated well with IL-8 and MCP-3 levels and monocyte activation. These changes were also reflected in miRNA profile in both groups.ConclusionsMV and miRNA could potentially serve as early AKI markers in children and their analysis reveals mechanistic parallels with adults.
IntroductionAllogeneic red blood cell (RBC) transfusion in cardiac surgery patients increases the risk of pulmonary and renal morbidity. These associations are attributable to the ‘storage lesion’ - ...accumulation of metabolites in the RBC supernatant that includes pro-inflammatory lipids, cell-free hemoglobin and energy depletion.HypothesisStorage lesion components trigger different events that lead to organ injury. Reducing storage lesion by RBC washing or rejuvenation will reduce organ injury.MethodsSamples from 60 cardiac patients participating in a randomized controlled trial of RBC washing prior to transfusion versus standard care were analysed for inflammatory markers. In a complementary experiment, 4 groups of pigs received crystalloid infusion or 14-day old RBC that were left untreated, washed or rejuvenated (Rejuvesol, Citra Labs, IN). Inflammatory markers, free hemoglobin and iron were measured in blood. Leukocyte invasion was determined by immunohistochemistry. The results were verified and further analysed in an in vitro inflammatory model.ResultsTransfusion of stored RBC (~22 days) into humans or pigs was associated with leukocyte activation, and attenuated by washing. That was attributed to oxidised lipids on transfused RBC derived microvesicles that activated monocytes and platelets, as revealed in depletion experiments. Unexpectedly, transfusion of washed RBC increased kidney injury, which correlated with higher free haemoglobin levels, oxidative stress (confirmed by HMOX-1 levels and protein carbonylation), and endothelial injury in washed RBC recipients. Oxidative stress triggered non-standard endothelial activation defined by expression of VLA5 and fibronectin retention. Either pathway was reduced when rejuvenated blood was transfused.ConclusionMonocytes/platelets and endothelium are activated by oxidized lipids/MV and reactive free haem, respectively, and can be alleviated by RBC rejuvenation.
Members of the Kinesin-3 family of kinesin-like proteins mediate transport of axonal vesicles (KIF1A, KIF1Bbeta), distribution of mitochondria (KIF1Balpha) and anterograde Golgi to ER vesicle ...transport (KIF1C). Until now, little is known about the regulation of kinesin-like proteins. Several proteins interact with members of this protein family. Here we report on a novel, KIF1 binding protein (KBP) that was identified in yeast two-hybrid screens.
KBP was identified by using the yeast-two-hybrid system with an amino-terminal fragment of KIF1C as a bait that is strongly homologous to KIF1B. Here we investigated the interaction of KBP and KIF1B. The full length proteins coimmunoprecipitated after overexpression and in untransfected 293 cells. Immunofluorescence experiments revealed that KBP was mainly localized to mitochondria, as has been described for KIF1Balpha. Overexpression of a deletion mutant or reduction of the KBP protein level using an anti-sense construct led to an aggregation of mitochondria. Such an effect is probably due to the lower activity of KIF1Balpha in the absence of KBP, as was revealed in motility assays.
KBP is a new binding partner for KIF1Balpha that is a regulator of its transport function and thus represents a new type of kinesin interacting protein.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Mycotoxins are secondary fungal metabolites, toxic to humans, animals and plants. Under the influence of various factors, mycotoxins may undergo modifications of their chemical structure. One of the ...methods of mycotoxin modification is a transformation occurring in plant cells or under the influence of fungal enzymes. This paper reviews the current knowledge on the natural occurrence of the most important trichothecenes and zearalenone in cereals/cereal products, their metabolism, and the potential toxicity of the metabolites. Only very limited data are available for the majority of the identified mycotoxins. Most studies concern biologically modified trichothecenes, mainly deoxynivalenol-3-glucoside, which is less toxic than its parent compound (deoxynivalenol). It is resistant to the digestion processes within the gastrointestinal tract and is not absorbed by the intestinal epithelium; however, it may be hydrolysed to free deoxynivalenol or deepoxy-deoxynivalenol by the intestinal microflora. Only one zearalenone derivative, zearalenone-14-glucoside, has been extensively studied. It appears to be more reactive than deoxynivalenol-3-glucoside. It may be readily hydrolysed to free zearalenone, and the carbonyl group in its molecule may be easily reduced to α/β-zearalenol and/or other unspecified metabolites. Other derivatives of deoxynivalenol and zearalenone are poorly characterised. Moreover, other derivatives such as glycosides of T-2 and HT-2 toxins have only recently been investigated; thus, the data related to their toxicological profile and occurrence are sporadic. The topics described in this study are crucial to ensure food and feed safety, which will be assisted by the provision of widespread access to such studies and obtained results.
Abstract
Agriculture plays a pivotal role in the economies of developing countries by providing livelihoods, sustenance, and employment opportunities in rural areas. However, crop diseases pose a ...significant threat to both farmers’ incomes and food security. Furthermore, these diseases also show adverse effects on human health by causing various illnesses. Till date, only a limited number of studies have been conducted to identify and classify diseased cauliflower plants but they also face certain challenges such as insufficient disease surveillance mechanisms, the lack of comprehensive datasets that are properly labelled as well as are of high quality, and the considerable computational resources that are necessary for conducting thorough analysis. In view of the aforementioned challenges, the primary objective of this manuscript is to tackle these significant concerns and enhance understanding regarding the significance of cauliflower disease identification and detection in rural agriculture through the use of advanced deep transfer learning techniques. The work is conducted on the four classes of cauliflower diseases i.e. Bacterial spot rot, Black rot, Downy Mildew, and No disease which are taken from VegNet dataset. Ten deep transfer learning models such as EfficientNetB0, Xception, EfficientNetB1, MobileNetV2, EfficientNetB2, DenseNet201, EfficientNetB3, InceptionResNetV2, EfficientNetB4, and ResNet152V2, are trained and examined on the basis of root mean square error, recall, precision, F1-score, accuracy, and loss. Remarkably, EfficientNetB1 achieved the highest validation accuracy (99.90%), lowest loss (0.16), and root mean square error (0.40) during experimentation. It has been observed that our research highlights the critical role of advanced CNN models in automating cauliflower disease detection and classification and such models can lead to robust applications for cauliflower disease management in agriculture, ultimately benefiting both farmers and consumers.
In these studies, we designed and investigated the potential anticancer activity of five iron(II) cyclopentadienyl complexes bearing different phosphine and phosphite ligands. All complexes were ...characterized with spectroscopic analysis viz. NMR, FT-IR, ESI-MS, UV-Vis, fluorescence, XRD (for four complexes) and elemental analyses. For biological studies, we used three types of cells-normal peripheral blood mononuclear (PBM) cells, leukemic HL-60 cells and non-small-cell lung cancer A549 cells. We evaluated cell viability and DNA damage after cell incubation with these complexes. We observed that all iron(II) complexes were more cytotoxic for HL-60 cells than for A549 cells. The complex CpFe(CO)(P(OPh)
)(η
-N-maleimidato) 3b was the most cytotoxic with IC
= 9.09 µM in HL-60 cells, IC
= 19.16 µM in A549 and IC
= 5.80 µM in PBM cells. The complex CpFe(CO)(P(Fu)
)(η
-N-maleimidato) 2b was cytotoxic only for both cancer cell lines, with IC
= 10.03 µM in HL-60 cells and IC
= 73.54 µM in A549 cells. We also found the genotoxic potential of the complex 2b in both types of cancer cells. However, the complex CpFe(CO)
(η
-N-maleimidato) 1 which we studied previously, was much more genotoxic than complex 2b, especially for A549 cells. The plasmid relaxation assay showed that iron(II) complexes do not induce strand breaks in fully paired ds-DNA. The DNA titration experiment showed no intercalation of complex 2b into DNA. Molecular docking revealed however that complexes CpFe(CO)(PPh
) (η
-N-maleimidato) 2a, 2b, 3b and CpFe(CO)(P(OiPr)
)(η
-N-maleimidato) 3c have the greatest potential to bind to mismatched DNA. Our studies demonstrated that the iron(II) complex 1 and 2b are the most interesting compounds in terms of selective cytotoxic action against cancer cells. However, the cellular mechanism of their anticancer activity requires further research.
Biodegradable 3D-printed polycaprolactone scaffolds for bone tissue engineering applications have been extensively studied as they can provide an attractive porous architecture mimicking natural ...bone, with tunable physical and mechanical properties enhancing positive cellular response. The main drawbacks of polycaprolactone-based scaffolds, limiting their applications in tissue engineering are: their hydrophobic nature, low bioactivity and poor mechanical properties compared to native bone tissue. To overcome these issues, the surface of scaffolds is usually modified and covered with a ceramic layer. However, a detailed description of the adhesion forces of ceramic particles to the polymer surface of the scaffolds is still lacking. Our present work is focused on obtaining PCL-based composite scaffolds to strengthen the architecture of the final product. In this manuscript, we report qualitative and quantitative evaluation of low temperature plasma modification followed by detailed studies of the adhesion forces between chemically attached ceramic layer and the surface of polycaprolactone-nanohydroxyapatite composite 3D-printed scaffolds. The results suggest modification-dependent alteration of the internal structure and morphology, as well as mechanical and physical scaffold properties recorded with atomic force microscopy. Moreover, changes in the material surface were followed by enhanced adhesion forces binding the ceramic layer to polymer-based scaffolds.
•Polycaprolactone-based bone tissue engineering scaffolds were fabricated and modified using low-temperature plasma.•Low temperature plasma modification of scaffolds altered their surface structure, roughness, stiffness and wettability.•The scaffolds were coated with hydroxyapatite and adhesion forces between the coating and polymeric surface was assesed.
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