Tick saliva contains a number of effector molecules that inhibit host immunity and facilitate pathogen transmission. How tick proteins regulate immune signaling, however, is incompletely understood. Here, we describe that loop 2 of sialostatin L2, an anti-inflammatory tick protein, binds to annexin A2 and impairs the formation of the NLRC4 inflammasome during infection with the rickettsial agent Anaplasma phagocytophilum Macrophages deficient in annexin A2 secreted significantly smaller amounts of interleukin-1β (IL-1β) and IL-18 and had a defect in NLRC4 inflammasome oligomerization and caspase-1 activation. Accordingly, Annexin a2-deficient mice were more susceptible to A. phagocytophilum infection and showed splenomegaly, thrombocytopenia, and monocytopenia. Providing translational support to our findings, better binding of annexin A2 to sialostatin L2 in sera from 21 out of 23 infected patients than in sera from control individuals was also demonstrated. Overall, we establish a unique mode of inflammasome evasion by a pathogen, centered on a blood-feeding arthropod.

• MeSH
• Anaplasma phagocytophilum genetics   immunology   MeSH
• Annexin A2 chemistry   genetics   immunology   MeSH
• Arachnid Vectors chemistry   genetics   immunology   MeSH
• Cystatins chemistry   genetics   immunology   MeSH
• Ehrlichiosis immunology   microbiology   pathology   MeSH
• Escherichia coli genetics   metabolism   MeSH
• Immune Evasion * MeSH
• Inflammasomes genetics   immunology   MeSH
• Interleukin-18 genetics   immunology   MeSH
• Interleukin-1beta genetics   immunology   MeSH
• Caspase 1 genetics   immunology   MeSH
• Caspases genetics   immunology   MeSH
• Ixodes chemistry   genetics   immunology   MeSH
• Humans MeSH
• Macrophages immunology   microbiology   MeSH
• Models, Molecular MeSH
• Mice MeSH
• Protein Isoforms chemistry   genetics   immunology   MeSH
• Apoptosis Regulatory Proteins chemistry   genetics   immunology   MeSH
• Calcium-Binding Proteins chemistry   genetics   immunology   MeSH
• Gene Expression Regulation MeSH
• Recombinant Proteins chemistry   genetics   immunology   MeSH
• Amino Acid Sequence MeSH
• Signal Transduction MeSH
• Protein Binding MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Ciel' ghansových bunkách, v jadrách endotelu fetálnych kapilár a v cytoplazme Hofbauerových buniek v obidvoch súboroch. Expresia v myofibroblastoch choriónovej strómy bola silnejšia v prípadoch preeklampsie. Expresia superoxiddismutázy dosahovala v materiáloch z preeklampsie slabšiu úroveň v syncýciu, Langhansových bunkách ako aj v bunkách deciduy. Kalretinín nevykázal expresiu v žiadnej štruktúre placenty. V bazálnej decidue z prípadov preeklampsie bol ojedinele prítomný v bunkách intersticiálneho extravilózneho trofoblastu, v deciduálnych bunkách a v špirálovitých artériách. Záver: Naše pozorovania sú morfologickým príspevkom do objasňovania molekulového pozadia etiopatogenézy preeklampsie a podporujú jej niektoré klinicko-biochemické rysy.

Objective: To determine new data related to the expression of caspase 1, superoxiddismutase and calretinin in the placenta and basal decidua in preeclampsia. Material and methods: Placental and basal decidua samples from 9 preeclamptic and 9 normotensive controls were analyzed using expressions of caspase 1, superoxiddismutase and calretinin assesed by immunohistochemistry. Results: Caspase 1 was expressed in placental syncythium in preeclampsia constantly, while in the control group the expression was weak or absent. In Langhans cells, in fetal sinusoidal capillary endothelia and in Hofbauer cells the expression was equal in both groups. Stronger expression was observed in stromal myofibroblasts in preeclampsia. In preeclampsia, expression of superoxiddismutase in syncythium, in Langhans cells and in decidual cells was weaker. Calretinin was not found in any placental structure. Sporadically, calretinin was expressed in the interstitial extravillous trophoblast cells, in decidual cells and in spiral arterioles in preeclampsia. Conclusion: The obtained morphological data correlating with some clinical and biochemical features contribute to understanding of the molecular background of preeclampsia etiopathogenesis.

• MeSH
• Decidua MeSH
• Financing, Organized MeSH
• Immunohistochemistry methods   utilization   MeSH
• Caspase 1 isolation & purification   MeSH
• Clinical Laboratory Techniques utilization   MeSH
• Humans MeSH
• Eye Proteins isolation & purification   MeSH
• Placenta MeSH
• Pre-Eclampsia diagnosis   MeSH
• Nerve Tissue Proteins isolation & purification   MeSH
• S100 Calcium Binding Protein G isolation & purification   MeSH
• Statistics as Topic MeSH
• Superoxide Dismutase isolation & purification   MeSH
• Check Tag
• Humans MeSH
• Female MeSH

Cytotoxicity and apoptosis induced by etoposide were studied during 72 hr in human melanoma cells. Etoposide initiated DNA-damage signaling via ATM kinase and activated p53 pathway and caspase-2. In response to treatment with etoposide, mitochondria of melanoma cells first increased their abundance and activity, and at later treatment intervals their dynamic behavior and functions became suppressed. Observed mitochondrial perturbation was not preceded by membrane potential loss but cytochrome c release was observed together with a rise in caspase-9 and caspase-3 activities. The pharmacological inhibition of relevant induced targets proved the importance of ATM and caspase-2 in etoposide-mediated cytotoxicity and apoptosis.

• MeSH
• Adenosine Triphosphate metabolism   MeSH
• Apoptosis drug effects   MeSH
• Benzothiazoles pharmacology   MeSH
• Cyclosporine pharmacology   MeSH
• Cysteine Endopeptidases metabolism   MeSH
• DNA-Binding Proteins antagonists & inhibitors   metabolism   MeSH
• Etoposide pharmacology   MeSH
• Antineoplastic Agents, Phytogenic pharmacology   MeSH
• Cysteine Proteinase Inhibitors pharmacology   MeSH
• Caspase Inhibitors MeSH
• Protein Kinase Inhibitors pharmacology   MeSH
• Caspase 2 metabolism   MeSH
• Caspase 3 metabolism   MeSH
• Caspase 9 metabolism   MeSH
• Humans MeSH
• Melanoma pathology   MeSH
• Mitochondria drug effects   MeSH
• Tumor Cells, Cultured drug effects   MeSH
• Neoplasm Proteins antagonists & inhibitors   metabolism   MeSH
• Tumor Suppressor Proteins antagonists & inhibitors   metabolism   MeSH
• Tumor Suppressor Protein p53 metabolism   MeSH
• DNA Damage MeSH
• bcl-2-Associated X Protein metabolism   MeSH
• Protein Serine-Threonine Kinases antagonists & inhibitors   metabolism   MeSH
• Cell Cycle Proteins metabolism   MeSH
• Superoxides metabolism   MeSH
• Toluene analogs & derivatives   pharmacology   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH

BACKGROUND: Fatty acid-induced apoptosis and ER stress of pancreatic β-cells contribute to the development of type 2 diabetes, however, the molecular mechanisms involved are unclear. AIMS: In this study we have tested the role of caspase-2 and suggested ER stress mediator JNK in saturated fatty acid-induced apoptosis of the human pancreatic β-cells NES2Y. RESULTS: We found that stearic acid at apoptosis-inducing concentration activated ER stress signaling pathways, i.e. IRE1α, PERK and ATF6 pathways, in NES2Y cells. During stearic acid-induced apoptosis, JNK inhibition did not decrease the rate of apoptosis nor the activation of caspase-8, -9, -7 and -2 and PARP cleavage. In addition, inhibition of JNK activity did not affect CHOP expression although it did decrease the induction of BiP expression after stearic acid treatment. Caspase-2 silencing had no effect on PARP as well as caspase-8, -9 and -7 cleavage and the induction of CHOP expression, however, it also decreased the induction of BiP expression. Surprisingly, caspase-2 silencing was accompanied by increased phosphorylation of c-Jun. CONCLUSIONS: We have demonstrated that caspase-2 as well as JNK are not key players in apoptosis induction by saturated fatty acids in human pancreatic β-cells NES2Y. However, they appear to be involved in the modulation of saturated fatty acid-induced ER stress signaling, probably by a mechanism independent of c-Jun phosphorylation.

• MeSH
• Apoptosis drug effects   MeSH
• Insulin-Secreting Cells cytology   metabolism   MeSH
• DNA-Binding Proteins genetics   metabolism   MeSH
• Phosphorylation MeSH
• JNK Mitogen-Activated Protein Kinases antagonists & inhibitors   metabolism   MeSH
• Caspase 2 chemistry   genetics   metabolism   MeSH
• Caspase 7 metabolism   MeSH
• Caspase 8 metabolism   MeSH
• Caspase 9 metabolism   MeSH
• Stearic Acids pharmacology   MeSH
• Humans MeSH
• RNA, Small Interfering metabolism   MeSH
• Poly(ADP-ribose) Polymerases metabolism   MeSH
• Heat-Shock Proteins metabolism   MeSH
• RNA Interference MeSH
• Signal Transduction drug effects   MeSH
• Endoplasmic Reticulum Stress drug effects   MeSH
• Activating Transcription Factor 6 metabolism   MeSH
• Transcription Factor CHOP metabolism   MeSH
• Transcription Factors genetics   metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The aim of this study was to characterize an in vitro modulating effect of three commensal Lactobacillus strains on cellular differentiation of non-transformed crypt-like rat small intestinal cell line IEC-18. IEC-18 was grown on extracellular matrix, with or without presence of Lactobacillus strains. Gene expression of IEC-18 bacterial detection system - such as Toll-like receptors TLR-2, TLR-4, signal adapter MyD88, cytoplasmic NOD2 receptor, inflammatory cytokines IL-18, IL-1beta, chemokine IL-8 and enzyme caspase-1 - was evaluated using real-time PCR. Expression and localization of TLR-2, TLR-4, IL-18 and caspase-1 proteins was demonstrated by Western blotting and immunofluorescent staining. Secretion of IL-18 to apical and basolateral surfaces was assayed by ELISA. Our results suggested that L. casei LOCK0919 accelerated differentiation of IEC-18 by stimulating TLR-2, TLR-4, MyD88, IL-18, caspase-1 mRNAs and proteins. L. casei LOCK0919 increased expression and transfer of villin and beta-catenin from cytoplasm to cell membrane. Presence of L. rhamnosus LOCK0900 resulted in detachment of IEC-18 layer from extracellular matrix leading to induction of IL-1beta, of TLR-2 and IL-8 mRNAs and stimulation of MyD88, caspase-1 and cytosolic receptor NOD2 mRNAs. L. rhamnosus LOCK0908 was not recognized by TLR-2 or TLR-4 receptors. Lactobacilli-IEC-18 crosstalk enhanced immune and barrier mucosal functions.

• MeSH
• beta Catenin biosynthesis   MeSH
• Cell Differentiation drug effects   MeSH
• Cytokines biosynthesis   MeSH
• Epithelial Cells drug effects   MeSH
• Interleukin-18 biosynthesis   MeSH
• Caspase 1 biosynthesis   MeSH
• Rats MeSH
• Lacticaseibacillus rhamnosus * MeSH
• Lacticaseibacillus casei * MeSH
• RNA, Messenger biosynthesis   MeSH
• Microfilament Proteins biosynthesis   MeSH
• Probiotics pharmacology   MeSH
• Gene Expression Regulation drug effects   MeSH
• Intestinal Mucosa cytology   drug effects   MeSH
• Subcellular Fractions metabolism   MeSH
• Toll-Like Receptors biosynthesis   drug effects   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Although EcoR124 is one of the better-studied Type I restriction-modification enzymes, it still presents many challenges to detailed analyses because of its structural and functional complexity and missing structural information. In all available structures of its motor subunit HsdR, responsible for DNA translocation and cleavage, a large part of the HsdR C terminus remains unresolved. The crystal structure of the C terminus of HsdR, obtained with a crystallization chaperone in the form of pHluorin fusion and refined to 2.45 Å, revealed that this part of the protein forms an independent domain with its own hydrophobic core and displays a unique α-helical fold. The full-length HsdR model, based on the WT structure and the C-terminal domain determined here, disclosed a proposed DNA-binding groove lined by positively charged residues. In vivo and in vitro assays with a C-terminal deletion mutant of HsdR supported the idea that this domain is involved in complex assembly and DNA binding. Conserved residues identified through sequence analysis of the C-terminal domain may play a key role in protein-protein and protein-DNA interactions. We conclude that the motor subunit of EcoR124 comprises five structural and functional domains, with the fifth, the C-terminal domain, revealing a unique fold characterized by four conserved motifs in the IC subfamily of Type I restriction-modification systems. In summary, the structural and biochemical results reported here support a model in which the C-terminal domain of the motor subunit HsdR of the endonuclease EcoR124 is involved in complex assembly and DNA binding.

• MeSH
• Biophysical Phenomena MeSH
• DNA-Binding Proteins chemistry   genetics   MeSH
• Escherichia coli chemistry   genetics   MeSH
• Protein Conformation MeSH
• Crystallography, X-Ray MeSH
• Multiprotein Complexes chemistry   genetics   MeSH
• Protein Subunits chemistry   genetics   MeSH
• Protein Domains genetics   MeSH
• Escherichia coli Proteins chemistry   genetics   MeSH
• Deoxyribonucleases, Type I Site-Specific chemistry   genetics   MeSH
• Amino Acid Sequence MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Caspase-3 and -7 are generally known for their central role in the execution of apoptosis. However, their function is not limited to apoptosis and under specific conditions activation has been linked to proliferation or differentiation of specialised cell types. In the present study, we followed the localisation of the activated form of caspase-7 during intramembranous (alveolar and mandibular bones) and endochondral (long bones of limbs) ossification in mice. In both bone types, the activated form of caspase-7 was detected from the beginning of ossification during embryonic development and persisted postnatally. The bone status was investigated by microCT in both wild-type and caspase-7-deficient adult mice. Intramembranous bone in mutant mice displayed a statistically significant decrease in volume while the mineral density was not altered. Conversely, endochondral bone showed constant volume but a significant decrease in mineral density in caspase-7 knock-out mice. Cleaved caspase-7 was present in a number of cells that did not show signs of apoptosis. PCR array analysis of the mandibular bone of caspase-7-deficient versus wild-type mice pointed to a significant decrease in mRNA levels for Msx1 and Smad1 in early bone formation. These observations might explain the decrease in the alveolar bone volume of adult knock-out mice. In conclusion, this study is the first to report a non-apoptotic function of caspase-7 in osteogenesis and also demonstrates further specificities in endochondral versus intramembranous ossification.

• MeSH
• Apoptosis MeSH
• Embryonic Development MeSH
• Caspase 3 metabolism   MeSH
• Caspase 7 genetics   metabolism   MeSH
• Bone and Bones metabolism   pathology   radiography   MeSH
• Mice, Knockout MeSH
• Mice MeSH
• Osteogenesis * MeSH
• Osteocalcin metabolism   MeSH
• Tomography, X-Ray Computed MeSH
• Smad1 Protein genetics   metabolism   MeSH
• MSX1 Transcription Factor genetics   metabolism   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Přirozená imunita hraje významnou roli v obraně proti infekci. Jednou z komponent přirozené humorální imunity je i manózu vázající lektin. Ten je velmi intenzivně studován již přes dvacet let. Tento článek shrnuje informace o základních charakteristikách manózu vázajícího lektinu a možnostech jeho laboratorního vyšetření. Zároveň přináší přehled klinických studií zkoumajících vztah manózu vázajícího lektinu k infekci. Výstupy klinických studií lze využít v imunologické praxi jak k indikaci laboratorního vyšetření manózu vázajícího lektinu, tak k interpretaci jeho výsledných hodnot.

The innate immunity plays a very important role in the defence against infection. Humoral section of the innate immunity also i ncludes manno- se-binding lectin. Mannose-binding lectin has been studied extensively for more than twenty years. This article summarizes bas ic facts about mannose-binding lectin including assays for its laboratory evaluation. At the same time it brings in a survey of clinical studi es researching mannose-binding lectin in relation to infection. Outcomes of the studies can be used in everyday practice, both in the indicati on of the laboratory evaluation of mannose-binding lectin and for its interpretation.

• Keywords
• MBL, komplementový systém,
• MeSH
• Child MeSH
• Infections diagnosis   MeSH
• Clinical Laboratory Techniques methods   utilization   MeSH
• Infant MeSH
• Complement System Proteins immunology   deficiency   MeSH
• Mannose-Binding Lectin analysis   diagnostic use   blood   MeSH
• Humans MeSH
• Infant, Newborn MeSH
• Child, Preschool MeSH
• Immunity, Innate physiology   genetics   immunology   MeSH
• Check Tag
• Child MeSH
• Infant MeSH
• Humans MeSH
• Infant, Newborn MeSH
• Child, Preschool MeSH

The cellular and molecular pharmacology of the new class of anticancer drugs, in which the CDK inhibitor bohemine and its analogues are coordinated to Pt(II) to form cisplatin derivatives, was investigated. The results revealed the unique anticancer profile of a cisplatin-derived platinum(II) dichlorido complex involving N(7)-coordinated bohemine (C1). Although the IC(50) values were ∼6-fold higher for C1 than for cisplatin in cisplatin-sensitive tumor cells, the tumor cells in which C1 was also active are those which acquired resistance to cisplatin. In addition, among the novel conjugates of bohemine and its analogues with cisplatin, marked selectivity of C1 for tumor cells relative to the nontumorigenic, normal cells was observed. However, coordination of bohemine to platinum in C1 considerably reduced one of the dual functionalities anticipated to be effective after C1 reaches the nucleus. Further studies performed in the cells with wt p53 status show differences between cisplatin and C1 at the level of cell cycle regulation. Impedance-based real-time monitoring of the effects of C1 and cisplatin on cell growth supported the thesis that critical differences exist in the rate and mechanisms of cell kill caused by the two agents and that C1 was a more potent inducer of apoptosis and/or necrosis than cisplatin. The results also showed that the distinct differences in cell killing observed for C1 and cisplatin might be associated with processes at the DNA level. The DNA binding experiments carried out in a cell-free medium demonstrated that modification reactions resulting in the irreversible coordination of C1 to DNA were slower than that of cisplatin. Transcription mapping experiments and determination of interstrand cross-linking efficiency of C1 suggested that several aspects of DNA binding mode of C1 and cisplatin were similar. It was concluded that C1 remains a promising prototype of compounds for the generation of novel drug candidates with cytotoxicity profiles different from those of the platinum drugs currently in use.

• MeSH
• Cell Line MeSH
• Cell Cycle drug effects   MeSH
• CHO Cells MeSH
• Cisplatin analogs & derivatives   pharmacology   MeSH
• Cricetulus MeSH
• DNA metabolism   MeSH
• Protein Kinase Inhibitors pharmacology   MeSH
• Cricetinae MeSH
• Humans MeSH
• Cell Line, Tumor MeSH
• CDC2 Protein Kinase antagonists & inhibitors   MeSH
• Antineoplastic Agents pharmacology   MeSH
• Purines pharmacology   MeSH
• Cell Survival drug effects   MeSH
• Animals MeSH
• Check Tag
• Cricetinae MeSH
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Changes in the levels of calcium binding proteins are known to occur in different parts of the brain during aging. In our study we attempted to define the effect that aging has on the parvalbumin-expressing system of neurons in the higher parts of the central auditory system. Age-related changes in parvalbumin immunoreactivity were investigated in the inferior colliculus (IC), medial geniculate body (MGB) and auditory cortex (AC) in two rat strains, normally aging Long-Evans (LE) and fast aging Fischer 344 (F344). The results demonstrate that the changes in PV-immunoreactivity are strain-dependent with an increase in the number of PV-immunoreactive (PV-ir) neurons occurring in the inferior colliculus of old LE rats and a pronounced decline in the number of PV-ir neurons appearing in the auditory cortex of aged F344 animals. In some parts of the AC of old F344 animals no PV-ir neurons were present at all. The number of PV-ir neurons in the MGB in all examined animals was very low independent of the strain and age. The loss of PV-ir neurons in the auditory cortex of Fischer 344 rats with aging may contribute to the substantial deterioration of hearing function in this strain.

• MeSH
• Inferior Colliculi metabolism   MeSH
• Species Specificity MeSH
• Financing, Organized MeSH
• Rats MeSH
• Geniculate Bodies metabolism   MeSH
• Neurons metabolism   MeSH
• Parvalbumins metabolism   MeSH
• Rats, Inbred F344 MeSH
• Rats, Long-Evans MeSH
• Calcium-Binding Proteins metabolism   MeSH
• Auditory Cortex metabolism   MeSH
• Aging metabolism   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Animals MeSH