Human Hsp40 proteins, DNAJA1 and DNAJA2, as potential targets of the immune response triggered by bacterial DnaJ in rheumatoid arthritis

Hsp40 proteins of bacterial and human origin are suspected to be involved in the pathogenesis of rheumatoid arthritis (RA). It has been shown that sera of RA patients contain increased levels of antibodies directed to bacterial and human Hsp40s. The aim of this work was to explore immunological similarities between the bacterial (DnaJ) and human (DNAJA1 and DNAJA2) Hsp40 proteins in relation to their possible involvement in the RA. Using polyclonal antibodies directed against a full-length DnaJ or its domains, against DNAJA1 and DNAJA2, as well as monoclonal anti-DnaJ antibodies, we found immunological similarities between the bacterial and human Hsp40s. Both ELISA and Western blotting showed that these similarities were not restricted to the conserved J domains but were also present in the C-terminal variable regions. We also found a positive correlation between the levels of the anti-DnaJ and anti-DNAJA1 antibodies in the sera of RA patients. This finding supports the molecular mimicry hypothesis that human Hsp40 could be the targets of antibodies originally directed against bacterial DnaJ in RA. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s12192-013-0407-1) contains supplementary material, which is available to authorized users

Education, medical care and treatment of women with various forms of high blood pressure during pregnancy

Nadciśnienie tętnicze jest jedną z najczęściej występujących patologii ciąży oraz najczęstszym czynnikiem śmiertelności matek w krajach rozwiniętych, a także źródłem wielu powikłań u dzieci. Pomimo ogromnego rozwoju wiedzy na temat etiopatologii i leczenia nadciśnienia w ciąży, jedynym przyczynowym leczeniem stanu przedrzucawkowego jest doprowadzenie do urodzenia dziecka i wydalenia łożyska. Aby w leczeniu i opiece nad ciężarną z nadciśnieniem osiągnąć zamierzony cel, należy uwzględnić nie tylko wiedzę i doświadczenie personelu medycznego, ale również współpracę pacjentki i jej świadome podejście do leczenia. Problemy Pielęgniarstwa 2010; 18 (4): 512-517Hypertension is one of the most common pathology in pregnancy and factors of maternal mortality in developed countries, as well as the source of many complications in children. Despite a significant progress in knowledge of etiopathology and treatment of high blood pressure during pregnancy the only causal treatment of preeclampsia is by induction of the labour of a child and placenta. Knowledge and experience of medical staff is of importance to achieve intended aims in treatment and care of hypertensive pregnant women; however, patient’s cooperation and an aware approach towards treatment is also essential. Nursing Topics 2010; 18 (4): 512-51

Care for pregnant women with diabetes

Cukrzyca jest najczęstszym powikłaniem metabolicznym ciąży. Przed odkryciem insuliny cukrzyca często uniemożliwiała donoszenie zdrowego dziecka. Od czasu wprowadzenia insulinoterapii rokowania dla przebiegu ciąży u kobiety z cukrzycą się poprawiły. Choroba ta ma jednak wpływ nie tylko na zdrowie i życie samych ciężarnych, ale także na różne objawy manifestowane przez noworodki matek chorych na cukrzycę. Ponadto stanowi również duże wyzwanie w opiece neonatologicznej. Wprowadzenie nowoczesnych metod nadzoru biofizycznego płodu w ostatnich latach ułatwiło ustalenie terminu ukończenia ciąży i pozwoliło w efekcie na zmniejszenie częstości zgonów donoszonych płodów. Jednocześnie w badaniach klinicznych wykazano, że wprowadzenie opieki nad kobietą przed zajściem w ciążę i podczas jej pierwszego trymestru znacząco zmniejsza częstość występowania wad wrodzonych u dzieci tych kobiet w porównaniu z dziećmi kobiet chorych na cukrzycę niepoddanych opiece przed zapłodnieniem. Ważne jest więc, aby objęcie opieką medyczną wszystkich kobiet ciężarnych z cukrzycą następowało jeszcze w okresie prekoncepcyjnym. Lekarze i położne zajmujący się opieką nad ciężarnymi powinni prowadzić wczesną diagnostykę w kierunku cukrzycy ciężarnych według najnowszych zaleceń medycznych, a pozostały personel ochrony zdrowia powinien się zajmować szeroko rozumianą edukacją kobiet chorych na cukrzycę, ukierunkowaną na normalizację glikemii i zapobieganie powikłaniom. Problemy Pielęgniarstwa 2010; 18 (3): 348-352Diabetes is the most common metabolic complication of pregnancy. Before the discovery of insulin, diabetes often made it impossible for a pregnant woman to carry a healthy baby to term. Since the introduction of insulin therapy the prognoses for pregnant women with diabetes have improved. However, the disease not only has an impact on the health and life of pregnant women but the variety of symptoms manifested by fetuses of diabetic mothers is also a significant challenge in neonatal care. The introduction of modern methods of biophysical monitoring of the fetus in the recent years has made it easier to determine the term of delivery and consequently has resulted in reducing the mortality rate of full-term fetuses. Simultaneously, clinical tests have revealed that care for a woman before she gets pregnant and during the first trimester, considerably decreases the frequency of congenital defects in children of such women compared to children of women suffering from diabetes not cared for before conception. It is thus important to give medical care to all pregnant women with diabetes already in the preconception period. Therefore doctors and midwives caring for pregnant women should perform early diagnostics of GDM based on the latest medical recommendations and other members of the healthcare team should provide broadly understood education of women suffering from diabetes, aimed at glycaemia normalization and prevention of complications. Nursing Topics 2010; 18 (3): 348-35

Cytokines of the Th1 and Th2 type in sera of rheumatoid arthritis patients; correlations with anti-Hsp40 immune response and diagnostic markers

Rheumatoid arthritis (RA) is a chronic, systemic inflammatory disease which affects approximately 1% of the population worldwide. Recent research on the role of heat shock proteins (Hsps) in RA development indicates that they may have pro- or anti-inflammatory effect, most probably via modulating cytokine secretion. We investigated type Th1 (INFγ, TNFα, IL-2) and type Th2 (IL-10, IL-6, IL-4) cytokine levels in sera of RA patients and healthy controls, using flow cytometric bead array assay, and searched for correlations between the cytokine levels and serum antibodies against bacterial (DnaJ) and human (Hdj1, Hdj2 and Hdj3) Hsp40 proteins, as well as clinical and laboratory parameters. The levels of all cytokines studied were significantly increased in RA patients; the highest increase relative to healthy controls (7-fold) was observed for IL-6 and its levels correlated positively with the antibodies directed to DnaJ and to the C-terminal domain of Hdj2, and with diagnostic parameters (DAS 28, Steinbrocker RTG criteria, ARA/7, ESR, TEN, SW and GH). INFγ levels correlated negatively with DAS 28, ESR, TEN and SW. No correlations were found for TNFα, IL-2 or IL-4. Our results support the hypothesis of Hsp40 involvement in RA as well as indicate that IL-6 serum level is a good marker of the RA activity

Structural basis of the interspecies interaction between the chaperone DnaK(Hsp70) and the co-chaperone GrpE of archaea and bacteria

Hsp70s are chaperone proteins that are conserved in evolution and present in all prokaryotic and eukaryotic organisms. In the archaea, which form a distinct kingdom, the Hsp70 chaperones have been found in some species only, including Methanosarcina mazei. Both the bacterial and archaeal Hsp70(DnaK) chaperones cooperate with a GrpE co-chaperone which stimulates the ATPase activity of the DnaK protein. It is currently believed that the archaeal Hsp70 system was obtained by the lateral transfer of chaperone genes from bacteria. Our previous finding that the DnaK and GrpE proteins of M. mazei can functionally cooperate with the Escherichia coli GrpE and DnaK supported this hypothesis. However, the cooperation was surprising, considering the very low identity of the GrpE proteins (26%) and the relatively low identity of the DnaK proteins (56%). The aim of this work was to investigate the molecular basis of the observed interspecies chaperone interaction. Infrared resolution-enhanced spectra of the M. mazei and E. coli DnaK proteins were almost identical, indicating high similarity of their secondary structures, however, some small differences in band position and in the intensity of amide I' band components were observed and discussed. Profiles of thermal denaturation of both proteins were similar, although they indicated a higher thermostability of the M. mazei DnaK compared to the E. coli DnaK. Electrophoresis under non-denaturing conditions demonstrated that purified DnaK and GrpE of E. coli and M. mazei formed mixed complexes. Protein modeling revealed high similarity of the 3-dimensional structures of the archaeal and bacterial DnaK and GrpE proteins

The DnaK chaperones from the archaeon Methanosarcina mazei and the bacterium Escherichia coli have different substrate specificities

Hsp70 (DnaK) is a highly conserved molecular chaperone present in bacteria, eukaryotes, and some archaea. In a previous work we demonstrated that DnaK from the archaeon Methanosarcina mazei (DnaKMm) and the DnaK from the bacterium Escherichia coli (DnaKEc) were functionally similar when assayed in vitro but DnaKMm failed to substitute for DnaKEc in vivo. Searching for the molecular basis of the observed DnaK species specificity we compared substrate binding by DnaKMm and DnaKEc. DnaKMm showed a lower affinity for the model peptide (a-CALLQSRLLS) compared to DnaKEc. Furthermore, it was unable to negatively regulate the E. coli σ32 transcription factor level under heat shock conditions and poorly bound purified σ32, which is a native substrate of DnaKEc. These observations taken together indicate differences in substrate specificity of archaeal and bacterial DnaKs. Structural modeling of DnaKMm showed some structural differences in the substrate-binding domains of DnaKMm and DnaKEc, which may be responsible, at least partially, for the differences in peptide binding. Size-exclusion chromatography and native gel electrophoresis revealed that DnaKMm was found preferably in high molecular mass oligomeric forms, contrary to DnaKEc. Oligomers of DnaKMm could be dissociated in the presence of ATP and a substrate (peptide) but not ADP, which may suggest that monomer is the active form of DnaKMm

Assessment of Anxiety and Depression in Polish Primary Parental Caregivers of Children with Cerebral Palsy Compared to a Control Group, as well as Identification of Selected Predictors

Background: Taking care of a child with Cerebral Palsy (CP) may be linked with adverse effects in the parents’ physical and mental health. The causes of anxiety and depression symptoms associated with childcare are still not fully understood. Aim: To assess the intensity of anxiety and depression symptoms in parents of children with CP compared to a control group and to identify selected mental health predictors. Design and Methods: Data were collected from 301 respondents, including 190 parents of children with CP (study group) and 111 parents taking care of children developing normally (control group). Intensity of anxiety and depression was rated using the Hospital Anxiety and Depression Scale (HADS) scale. Gross Motor Function Classification System for Cerebral Palsy (GMFCS), Sense of Coherence Scale (SOC-29), Berlin Support Social Scales (BSSS) scales and a specially designed questionnaire were used to assess the predictors. The investigated variables included the children’s and the parents’ characteristics, as well as environmental factors. The analyses applied Spearman’s rank correlation coefficient, M(SD) as well as multiple regression. Results: The level of anxiety and depression was clearly higher in the parents of children with CP–the mean levels of anxiety and depression in the study group and the controls amounted to 8.1 vs. 4.7 and 6.8 vs. 3.7, respectively. The factors associated with intensity of anxiety and depression in the parents of children with CP included lack of social support, mainly perceived and received support, unsatisfying parental health status, poor economic status of the family, as well as difficult living conditions, sense of coherence, loneliness, the parent’s gender, and the child’s intellectual disability. Conclusions: Identification of significant anxiety and depression predictors, understood as modifiable factors, should be considered in determining and planning comprehensive support for a child with CP and his/her primary parental caregiver

Characterization of the anti-DnaJ monoclonal antibodies and their use to compare immunological properties of DnaJ and its human homologue HDJ-1

Escherichia coli DnaJ (Hsp40) is suspected to participate in rheumatoid arthritis (RA) pathogenesis in humans by an autoimmune process. In this work a set of 6 anti-DnaJ monoclonal antibodies (mAbs) was raised and localization of the epitopes recognized by the mAbs was investigated. Western blotting and enzyme-linked immunosorbent assay (ELISA) experiments showed that the mAbs efficiently bound only native antigen. Using DnaJ mutant proteins with deletions of specified domains and ELISA, we found that AC11 mAb reacted with the best conserved in evolution N-terminal J domain, whereas BB3, EE11, CC5, CC8, and DC7 bound to the C-terminal part after residue 200. Mapping performed with the use of a random peptide library displayed by filamentous phage indicated that (1) AC11 mAb bound to a region between residues 33–48, including D-34 which belongs to the HPD triad, present in all DnaJ homologues, (2) BB3 recognized residues localized in the 204–224 region, (3) EE11 recognized the 291–309 region, (4) CC5—the region 326–359, and (5) CC8—the 346–366 region. All these mAbs, as well as the polyclonal antibodies against the N- or C-terminal domain, bound efficiently to HDJ-1, human Hsp40. These results show the presence of a significant immunological similarity between bacterial DnaJ and human HDJ-1, which is not restricted to the evolutionarily conserved parts of the proteins, and suggest that HDJ-1 could be a possible target of immune response triggered by DnaJ