A roadmap for the conservation of freshwater mussels in Europe.

Europe has a long history of human pressure on freshwater ecosystems. As pressure continues to grow and new threats emerge, there is an urgent need for conservation of freshwater biodiversity and its ecosystem services. However, whilst some taxonomic groups, mainly vertebrates, have received a disproportionate amount of attention and funds, other groups remain largely off the public and scientific radar. Freshwater mussels (Bivalvia, Unionida) are an alarming example of this conservation bias and here we point out six conceptual areas that need immediate and long-term attention: knowledge, threats, socioeconomics, conservation, governance and education. The proposed roadmap aims to advance research, policy and education by identifying the most pressing priorities for the short- and long-term conservation of freshwater mussels across Europe.

Playing Peekaboo with a Master Manipulator: Metagenetic Detection and Phylogenetic Analysis of Wolbachia Supergroups in Freshwater Invertebrates.

The infamous "master manipulators"-intracellular bacteria of the genus Wolbachia-infect a broad range of phylogenetically diverse invertebrate hosts in terrestrial ecosystems. Wolbachia has an important impact on the ecology and evolution of their host with documented effects including induced parthenogenesis, male killing, feminization, and cytoplasmic incompatibility. Nonetheless, data on Wolbachia infections in non-terrestrial invertebrates are scarce. Sampling bias and methodological limitations are some of the reasons limiting the detection of these bacteria in aquatic organisms. In this study, we present a new metagenetic method for detecting the co-occurrence of different Wolbachia strains in freshwater invertebrates host species, i.e., freshwater Arthropoda (Crustacea), Mollusca (Bivalvia), and water bears (Tardigrada) by applying NGS primers designed by us and a Python script that allows the identification of Wolbachia target sequences from the microbiome communities. We also compare the results obtained using the commonly applied NGS primers and the Sanger sequencing approach. Finally, we describe three supergroups of Wolbachia: (i) a new supergroup V identified in Crustacea and Bivalvia hosts; (ii) supergroup A identified in Crustacea, Bivalvia, and Eutardigrada hosts, and (iii) supergroup E infection in the Crustacea host microbiome community.

Effects of extreme experimental drought and rewetting on CO2 and CH4 exchange in mesocosms of 14 European peatlands with different nitrogen and sulfur deposition.

The quantitative impact of intense drought and rewetting on gas exchange in ombrotrophic bogs is still uncertain. In particular, we lack studies investigating multitudes of sites with different soil properties and nitrogen (N) and sulfur (S) deposition under consistent environmental conditions. We explored the timing and magnitude of change in CO2 (Respiration, Gross Primary Production - GPP, and Net Exchange - NE) and CH4 fluxes during an initial wet, a prolonged dry (~100 days), and a subsequent wet period (~230 days) at 12 °C in 14 Sphagnum peat mesocosms collected in hollows from bogs in the UK, Ireland, Poland, and Slovakia. The relationship of N and S deposition with GPP, respiration, and CH4 exchange was investigated. Nitrogen deposition increased CO2 fluxes and GPP more than respiration, at least up to about 15 kg N ha(-1)  yr(-1) . All mesocosms became CO2 sources during drying and most of them when the entire annual period was considered. Response of GPP to drying was faster than that of respiration and contributed more to the change in NE; the effect was persistent and few sites recovered "predry" GPP by the end of the wet phase. Respiration was higher during the dry phase, but did not keep increasing as WT kept falling and peaked within the initial 33 days of drying; the change was larger when differences in humification with depth were small. CH4 fluxes strongly peaked during early drought and water table decline. After rewetting, methanogenesis recovered faster in dense peats, but CH4 fluxes remained low for several months, especially in peats with higher inorganic reduced sulfur content, where sulfate was generated and methanogenesis remained suppressed. Based on a range of European sites, the results support the idea that N and S deposition and intense drought can substantially affect greenhouse gas exchange on the annual scale.

Unusual manifestation of crystalline light chain tubulopathy in patient with multiple myeloma: case report and review of the literature.

Multiple myeloma (MM) is the second most common hematological malignancy, with an annual incidence in Europe and the USA of about 4-6 cases per 100,000. Several forms of renal disease are found in the course of MM, including: cast nephropathy, light chain (LC) deposition disease and primary amyloidosis. Less frequent forms include: acute and chronic tubulopathies, neoplastic plasma cell infiltration and interstitial nephritis. In this paper, we discuss a case of 53-year-old male patient with MM who presented with massive proteinuria (24 g/24 h), mild renal insufficiency (eGFR 43 mL/min), and Fanconi-like syndrome (as reflected by normoglycemic glycosuria). In kidney biopsy glomeruli were normal, whereas abundant AFOG-positive deposits were found in the cytoplasm of proximal tubular epithelial cells. These deposits were strongly positive for kappa light chains on immunofluorescence. Electron microscopy revealed electron-dense, intracytoplasmic crystalloid deposits of variable shape (needle-shaped, round and rectangular), and size in the proximal tubular cells. This unusual variant of microscopic renal lesions in the course of MM coupled with coincidence of Fanconi-like and nephrotic syndrome as a clinical manifestation has not been reported to date.

New paramedic scope of practice in Poland based on the 2022 reform.

Emergency medical systems in the world are mainly based on two main models: the Franco-German System (FGS) and the Anglo-American System (AAS). The characteristic feature of the FGS is the "Stay and Play" principle, while the AAS system is based on the "Scoop and Run" principle. The Polish model is a mix of those two systems mainly based on the work of paramedics. Their scope of operations and powers have changed over time. As a result of the advocacy undertaken by paramedics in Poland, legislation was drafted and became law in June 2023. The central changes include: the introduction of a paramedic register, the establishment of a professional self-government of paramedics, the expansion of professional competencies such as the ability to declare death, and new opportunities for professional development including speciality training or paid training leave. This article discusses the new law in the context of previous legislative solutions in the field of emergency medicine in Poland and in other European countries.

Is There Still a Place for Forceps Delivery in Modern Obstetrics?

BACKGROUND: Nowadays, we are witnessing a decrease of vaginal instrumental deliveries and continuous increase of caesarean section rate. However, proper identification of possibility of execution, indications for instrumental delivery and their skilful use may improve the broadly understood maternal and neonatal outcomes. The aim of this study is to present prevalence, risk factors, indications and outcomes of forceps deliveries among the patients at Department of Perinatology, Lodz. MATERIAL AND METHODS: A retrospective study was conducted at the Department of Perinatology, Medical University of Lodz. The study included forceps deliveries carried out between January 2019 and December 2022. Total number of 147 cases were analysed in terms of indications for forceps delivery and maternal and neonatal outcomes such as vaginal - or cervical - laceration, postpartum haemorrhage, perineal tear, newborn injuries, Apgar score, umbilical cord blood gas analysis, NICU admission and cranial ultrasound scans. RESULTS: The prevalence of forceps delivery was 2.2%. The most common indication for forceps delivery was foetal distress (81.6%). Among mothers, the most frequent complication was vaginal laceration (40.1%). Third-and fourth-degree perineal tears were not noted. Regarding neonatal outcomes, Apgar score ≥ 8 after 1st and 5th minute of life received accordingly 91.2% and 98% of newborns. Only 8.8% experienced severe birth injuries (subperiosteal haematoma, clavicle fracture). CONCLUSIONS: Although foetal distress is the most common indication for forceps delivery, the vast majority of newborns were born in good condition and did not require admission to NICU. Taking into consideration high efficacy and low risk of neonatal and maternal complications, forceps should remain in modern obstetrics.

Associations of Hormonal and Metabolic Parameters with Bone Mineralization in Young Adult Females.

Osteoporosis is characterized by impaired bone mineralization and microarchitecture. An important protective factor is a high peak bone mass (PBM), attained in the second and third decade of life. The aim of the study was to evaluate the effect of hormonal and metabolic parameters on bone mineralization in young adult female patients. A total of 111 participants qualified for the study. Bone mineral density of the lumbar spine (L1-L4) and whole skeleton was measured using dual-energy X-ray absorptiometry (DXA). Hormonal parameters were determined: the concentrations of androstendione, dihydroepiandrosterone sulphate, testosterone, sex hormone binding protein, 17-OH-progesterone, folliculotropic hormone, estradiol, thyrotropic hormone, free thyroxine and cortisol. Metabolic parameters were also examined. The study showed a statistically significant correlation between bone mineral density and estradiol concentration and a negative relationship between cortisol concentration and the bone mineral density (BMD) Z-score of the lumbar spine. Sclerostin measurements taken during this study were not related to bone mineral density. It has been shown that the concentration of the hormones tested, even within the reference range, may affect bone mineralization. We suggest observing the follow-up of the menstrual cycles, as well as analyzing the results of test patients in an annual examination system. However, each clinical case should be considered individually. The sclerostin test is currently not useful in the clinical evaluation of bone mineralization in young adult women.

Influence of Muscle Mass and Strength on Bone Mineralisation with Consideration of Sclerostin Concentration.

Osteoporosis is a disease characterised by a reduction in bone strength due to increased porosity and impaired mineralisation. In our study, we investigated whether muscle strength and mass exert a significant effect on bone mineral density in young adult women. We also tested whether sclerostin can be used as an indicator in the assessment of bone mineralisation. The study included 111 patients. All patients had their bone mineral density determined in the L1-L4 section of the lumbar spine and in the whole skeleton. The parameters of fat mass (FM), lean body mass (LBM) and visceral fat mass (VF) were also determined. Metabolic activity of osteocytes was assessed by measuring the serum sclerostin concentration. There was a statistically significant association of both hands' muscle strength with all parameters expressing bone mineralisation. A statistically significant relationship was also obtained between BMD L1-L4 and the body mass components (FM, LBM). Sclerostin levels in the study did not differ between groups with normal and reduced bone mineral density. Muscle strength assessment may be a potential exponent of reduced bone mineral density, also used clinically in young adult women. The utility of sclerostin in the clinical assessment of bone mineralisation has not been demonstrated.

Remodeling of Retinal Arterioles and Carotid Arteries in Heart Failure Development-A Preliminary Study.

Current data indicate that heart failure (HF) is associated with inflammation and microvascular dysfunction and remodeling. These mechanisms could be involved in HF development and progression, especially in HF with preserved ejection fraction (HFpEF). We aimed to compare structural changes in retinal arterioles and carotid arteries between HF patients and patients without heart failure. This preliminary, retrospective, case-control study included 28 participants (14 patients with HFpEF and 14 age- and sex-matched healthy controls). Carotid intima-media thickness to lumen ratio (cIMTLR) was assessed using B-mode ultrasonography. Retinal arterioles wall- to-lumen ratio (rWLR) was assessed by adaptive optics camera rtx1. The HF patients had higher IMTLR (Δmedian [HFpEF-control group] 0.07, p = 0.01) and eWLR (Δmedian 0.03, p = 0.001) in comparison to patients without HF. In the whole study group, rWLR correlated significantly with IMTLR (r = 0.739, p = 0.001). Prevalence of arterial hypertension was similar in both groups, however, patients with HF had a significantly lower office, central and 24-h ambulatory blood pressure (systolic Δmedian -21 to -18 mmHg; diastolic Δmedian -23 to -10 mmHg). Our data suggests gradual and simultaneous progression of vascular remodeling in both retinal arterioles and carotid arteries in HFpEF patients. This process could be a marker of HF development. Significantly lower blood pressure values in HF group may indicate that vascular remodeling could be independent of BP control. Nevertheless, further and larger prospective studies allowing to reduce the impact of confounding and address temporality are warranted.

Epicutaneous immunization with protein antigen in the presence of TLR4 ligand induces TCR alpha beta+CD4+ T contrasuppressor cells that reverse skin-induced suppression of Th1-mediated contact sensitivity.

Our previous work showed that epicutaneous (EC) immunization of mice with different protein Ags applied on the skin in the form of a patch induces a state of subsequent Ag-nonspecific unresponsiveness due to suppressor CD4+8+ T cells (Ts) that inhibit Th1-mediated contact sensitivity (CS) reactions via released TGF-beta. In the present work we show that EC immunization with Ag together with the TLR4 ligand LPS induced cells that could prevent suppression by the Ag-nonspecific Ts. These up-regulatory cells, called contrasuppressor T cells (Tcs), belong to a population of Ag-specific TCRalphabeta CD4+ lymphocytes and are different from Th1 CD4+ cells that mediate the CS reaction. Experiments using knockout mice showed that EC induced contrasuppression is MyD88, INF-gamma, and IL-12 dependent, whereas IL-6 is not involved in this phenomenon. Additional experiments with anti-IFN-gamma mAb showed that IFN-gamma is required for induction of Tcs cells but does not play a crucial role in the effector phase of contrasuppression. Additionally, treatment of CS effector cells with rIL-12 makes them resistant to EC induced suppression without affecting Ts cells, whereas IL-12 neutralization in vitro abrogates contrasuppression. These data show that IL-12 is indeed involved in the effector phase of EC induced contrasuppression and that this cytokine does not act directly on Ts cells. The mechanism of action of Tcs protects Th1 effector cells mediating CS from the nonspecific Ts, leaving suppression to other Ags intact. Ts and Tcs cells do not influence each other and can be induced simultaneously in the same animal.

Growth patterns of the pan-European freshwater mussel, Anodonta anatina (Linnaeus, 1758) (Bivalvia: Unionidae), vary with sex and mortality in populations.

Post-maturation growth leading to indeterminate growth patterns is widespread in nature. However, its adaptive value is unclear. Life history theory suggests this allocation strategy may be favored by temporal pulses in the intensity of mortality and/or the capacity to produce new tissues.Addressing the origin of indeterminate growth and the variability of growth patterns, we studied the growth of duck mussels, Anodonta anatina, a pan-European unionid, in 18 Polish lakes. For each population, the sex, size, and age of collected mussels were measured to estimate Bertalanffy's growth curve parameters. We integrated information on A. anatina mortality rates, lake trophy, biofouling by zebra mussels, Dreissena polymorpha, and the prevalence of parasitic trematode larvae to identify selective conditions in lakes.We found two sources of mortality in A. anatina populations, pertaining to adverse effects of zebra mussel biofouling and trophy state on mussel survival. Additionally, populations with heavier biofouling presented a smaller abundance of parasites, indicative of a relationship between filtering intensity and contraction of water-borne trematode larvae by filtering A. anatina.Consistently for each sex, populations with a greater trophy-related mortality were characterized in A. anatina by a smaller asymptotic size Lmax , indicative of a life history response to mortality risk involving early maturation at a smaller body size. In all populations, females featured higher mortality and larger asymptotic size versus males.Our findings support a theoretical view that adaptive responses to selection involve adjustments in the lifetime resource allocation patterns. These adjustments should be considered drivers of the origin of indeterminate growth strategy in species taking parental care by offspring brooding in body cavities.

The size and shape of parasitic larvae of naiads (Unionidae) are not dependent on female size.

The naiads, large freshwater mussels (Unionida), have very long life spans, are large-bodied, and produce thousands to millions of larvae (glochidia) which typically must attach to host fish tissues to metamorphose into a juvenile mussel. Glochidia develop within a female's marsupial gill demibranch, thus their number is restricted by female size. However, larger mussels acquire more energy, which could be invested in either larger-sized glochidia, in a more glochidia, or a combination of both. The high level of host specialization seen in many naiads may constrain glochidial size and shape around a narrow optimum, while naiads that use a wide range of host fishes may be predicted to possess greater plasticity in glochidial morphology. In this paper, we investigated the relationship between maternal body size and progeny body size and shape, aided by modern digital microscopy. We analyzed the between- and within- species variation of glochidia size and shape relative to female size in four widespread species of European naiads: Anodonta anatina, Anodonta cygnea, Unio crassus and Unio tumidus. Whereas the total reproductive output is collinear with female body size, substantial differences between species in glochidia size were found within genus Anodonta, but not genus Unio where glochidial size is remarkably consistent. The glochidial shape, however, differed within both Unio and Anodonta. We interpret this constant within-species glochidial size in Unio as reflecting a constraint imposed by the likelihood of successful transmission onto and off from a narrow range of hosts, whereas their shape seems to be less constrained. The Anodonta species, inhabiting a wide spectrum of habitats and using more than twice the number of fish hosts than Unio spp., have larger glochidia with greater variation in size and shape. Our results suggest that measures of glochidial variability may also serve as an indicator of host specificity in other naiads.

Response of freshwater mussel recruitment to hydrological changes in a eutrophic floodplain lake.

Although eutrophication of freshwaters is a natural process, the human impact often leads to inland waters becoming overloaded with nutrients, impoverishing many valuable and vanishing habitats, such as floodplain lakes. These changes need to be reversed if the occurrence of endangered aquatic species is to be restored. In this paper we analyse the impact of a change in the water regime of a naturally eutrophic floodplain lake, which harbours a large diversity of Unionidae (large freshwater mussels), a globally threatened taxonomic group that provides important ecosystem functions and services. We found that a slight increase in the discharge from this waterbody, following the construction of an additional outflow pipe, positively influenced recruitment in three of the five mussel species inhabiting the lake. We also found that, after the construction of this additional outflow, the niches of juveniles of Anodonta cygnea and Unio spp. changed, revealing differences in their hydrological requirements. Our results suggest that, as in lotic habitats, complex hydraulic parameters are highly significant to unionid mussels in lentic conditions.

The Patterns and Puzzles of Genetic Diversity of Endangered Freshwater Mussel Unio crassus Philipsson, 1788 Populations from Vistula and Neman Drainages (Eastern Central Europe).

Mussels of the family Unionidae are important components of freshwater ecosystems. Alarmingly, the International Union for Conservation of Nature and Natural Resources Red List of Threatened Species identifies almost 200 unionid species as extinct, endangered, or threatened. Their decline is the result of human impact on freshwater habitats, and the decrease of host fish populations. The Thick Shelled River Mussel Unio crassus Philipsson, 1788 is one of the examples that has been reported to show a dramatic decline of populations. Hierarchical organization of riverine systems is supposed to reflect the genetic structure of populations inhabiting them. The main goal of this study was an assessment of the U. crassus genetic diversity in river ecosystems using hierarchical analysis. Different molecular markers, the nuclear ribosomal internal transcribed spacer ITS region, and mitochondrial DNA genes (cox1 and ndh1), were used to examine the distribution of U. crassus among-population genetic variation at multiple spatial scales (within rivers, among rivers within drainages, and between drainages of the Neman and Vistula rivers). We found high genetic structure between both drainages suggesting that in the case of the analyzed U. crassus populations we were dealing with at least two different genetic units. Only about 4% of the mtDNA variation was due to differences among populations within drainages. However, comparison of population differentiation within drainages for mtDNA also showed some genetic structure among populations within the Vistula drainage. Only one haplotype was shared among all Polish populations whereas the remainder were unique for each population despite the hydrological connection. Interestingly, some haplotypes were present in both drainages. In the case of U. crassus populations under study, the Mantel test revealed a relatively strong relationship between genetic and geographical distances. However, in detail, the pattern of genetic diversity seems to be much more complicated. Therefore, we suggest that the observed pattern of U. crassus genetic diversity distribution is shaped by both historical and current factors i.e. different routes of post glacial colonization and history of drainage systems, historical gene flow, and more recent habitat fragmentation due to anthropogenic factors.

Bioactive yet antimicrobial structurally stable collagen/chitosan/lysine functionalized hyaluronic acid - based injectable hydrogels for potential bone tissue engineering applications.

Novel, biocompatible, multifunctional, injectable genipin crosslinked collagen/chitosan/lysine-modified hyaluronic acid based hydrogels (ColChHAmod) were prepared in a facile, one-step procedure. The novelty of the current approach lies in the functionalization of hyaluronic acid (HA) with primary amine groups by lysine attachment, and its further use as a component of the injectable sol. The obtained derivative, HAmod, could form, upon crosslinking with genipin, covalent bonds with other components of the hydrogel network, resulting in structurally stable, better-defined hydrogels. We have demonstrated that, by adjusting HAmod content and genipin concentration, hydrogels with tunable physicochemical characteristics (swelling, wettability, tendency for enzymatic degradation) and properties adequate for the potential bone tissue regeneration can be prepared. Storage modulus measurements indicated that HAmod has positive effect on mechanical characteristics of hydrogels prepared. It was also revealed that the ColChHAmod-based hydrogels are characterized by a high porosity (85-95%). The in situ rheological measurements confirmed the injectability of the obtained hydrogels. The in vitro cell culture studies showed that the surface of all materials prepared was biocompatible, as they supported proliferation and adhesion of osteoblast-like cells followed by ALP expression. The intrinsic antibacterial activity of the hydrogels against Escherichia coli was also demonstrated in in vitro experiment.

Role of TLR ligands in epicutaneously induced contrasuppression.

Our previous work showed that epicutaneous (EC) immunization in mice with protein antigen (Ag) induced an Ag-independent unresponsiveness mediated by suppressor CD4(+)8(+) T cells (Ts), which inhibited contact hypersensitivity (CS). Simultaneous EC immunization with Ag and various Toll-like receptor (TLR) ligands reversed skin-induced suppression. Our present study shows that this process activates Ag-specific T contrasuppressor (Tcs) cells and leads to the protection of CS effector T cells from suppression. Epicutaneous immunization with Ag and the TLR4 ligand lipopolysaccharide (LPS) led to a significant increase in IFN-gamma production by lymph node and spleen cells. Ag and TLR ligands, like LPS, CpG or lipoteichoic acid did not need to be applied concomitantly to the skin. An identical contrasuppressive effect was observed when the Ag and TLR ligands were deposited on distant skin areas, suggesting that both the generation of Ts and Tcs are independent. To corroborate this finding, we used a model system that uses macrophages (Mf) as Ag-presenting cells. Mf labeled in vitro with Ag (Mf-Ag) induced, upon intravenous (iv) administration, an unresponsiveness reaction that was mediated by Ts cells. When treated simultaneously with LPS-treated Mf (Mf-Ag-LPS), a TLR-ligand could induce CS. Both the Ag and the LPS signal could be uncoupled i.e., Mf-Ag and Mf-LPS given at separate time points (with an 1 h interval between injections) induced immunity.We also found that LPS-treated Mf also produced significant amounts of IL-12, a cytokine that has well-known anti-tolerogenic properties. Our experiments suggest that reversal of EC-induced suppression by TLR-ligands may be a potential tool to increase the immunogenicity of weakly immunogenic antigens.

TCRalphabeta + CD8+ and TCRgammadelta+ lymphocytes inhibit the capability of peritoneal macrophages to induce contact hypersensitivity.

Macrophages (Mf) play an important role in induction and regulation of the immune response. It was shown previously that subcutaneous injection of hapten conjugated macrophages (TNP-Mf) induces the contact hypersensitivity (CHS) response, whereas intravenous (i.v.) or intraperitioneal administration of TNP-Mf results in unresponsiveness as a result of induced T suppressor (Ts) cells. The aim of this study was to determine if different T cell populations influence macrophages to become inducers of immunological suppression. Our findings show that indeed i.v. injection of TNP labeled macrophages isolated from control mice into syngenic recipients induces unresponsiveness. However, i.v. administration of TNP substituted macrophages isolated from TCRalpha-/-, TCRdelta-/- and beta2m-/- mice induces strong CHS similar to that observed after skin painting with TNP-C1. Moreover, it was shown that TNP conjugated macrophages isolated from CD1d-/- mice were still able to promote immunosuppression when injected intravenously. This suggests that TCRalphabeta+ CD8+ and TCRgammadelta+ lymphocytes stimulate macrophages to induce immunosuppression instead of a strong CHS reaction, whereas CD1d dependent NKT cells are not involved in negative regulation of macrophage function.

Selective generation of lithiated benzonitriles: the importance of reaction conditions.

Lithiated benzonitriles can be generated in high yields from reactions of bromobenzonitriles with n-BuLi in THF under standard cryogenic conditions (ca. -70 degrees C) provided the reverse addition mode is employed. The resultant aryllithiums are fairly stable at temperatures up to -60 degrees C. The formation of lithiated benzonitriles via Br/Li exchange under normal addition mode conditions is plagued by deprotonation and extensive CN-addition reactions. The generation of related aryllithiums from disilylated bromobenzonitriles is comparatively less sensitive to reaction conditions.

Heme oxygenase (HO-1) is involved in the negative regulation of contact sensitivity reaction.

Cutaneous contact sensitivity (CS) is a subtype of delayed-type sensitivity and is mediated by either CD4(+) or CD8(+) CS-effector T cells. CS can be induced by skin painting with haptens like trinitrophenyl chloride (TNP-Cl).We have previously shown that CS is under the negative regulation of T regulatory cells (Treg) induced by the iv injection of a high dose of homologous antigen or via epicutaneous application of any protein antigen prior to TNP-Cl painting. In this study, we examined the role of heme oxygenase (HO-1) in the negative regulation of CS in mice. We found that ip injection of heme, an inducer of HO-1, before TNP-Cl sensitization strongly suppresses CS when compared to uninjected controls. Using a transfer out protocol, we showed that suppressor activity can be transferred with lymph node and spleen cells isolated from mice treated with heme for 7 days before TNP-Cl or sham immunization, which suggests a lack of antigen specificity of observed suppression. Negative selection with monoclonal antibodies and complement showed that regulatory cells induced via heme injection belong to the population of TCRalphabeta+ lymphocytes. Using CBA/J (H-2(k)), SJL (H-2(s)), and DBA1 (H-2(q)) mice, we showed that the suppression mediated by HO-1 is major histocompatibility complex (MHC) unrestricted. In vitro treatment of heme induced Treg cells with tin protoporphyrin IX (SnPPIX), an inhibitor of HO activity, prior to adoptive transfer abolished the suppressor activity. In summary, injection of heme results in the induction of antigen non-specific and MHC unrestricted TCRalphabeta+ Treg that suppress CS response in mice, possibly in a HO-1-dependent manner.

On the reintroduction of the endangered thick-shelled river mussel Unio crassus: The importance of the river's longitudinal profile.

Freshwater mussels of the order Unionida provide important ecosystem functions and services. Unfortunately, some previously widespread species are now seriously endangered. To restore the historical range of the population of Unio crassus in the Biala River, southern Poland, the species was reintroduced into a series of 'stepping stones' joining two remnant populations. During the first phase of the study, the relationships between the abundance of U. crassus, physical habitat, and water quality were studied to assess reintroduction potential. In general, chemical water quality improved upstream from the existing population, favouring the decision for reintroduction, whereas morphological variables worsened. Mussel abundance was correlated negatively with the elevation and slope of channel, organic matter contents, and pH (exceeding 8.0), but positively with silt presence, water conductivity, and concentrations of HCO3-, Ca2+, and NO3-. During the second phase, adult individuals were introduced into one type of functional habitat-marginal channel sectors with still water and fine sediment. Despite the initial very high rate of reproduction in some parts of the upper reach of the river, the juveniles were ultimately recruited only in the lower part of the restored range, resulting in a very rapid change in recruitment at a channel slope of 1.8‰. Recruitment was positively related to silt content, conductivity, and Ca2+ and HCO3- ions, negatively to channel elevation and slope, and water pH. The host fish species showed no correlation with abiotic habitat features within the studied reach. These results imply that most of the habitat traits related to U. crassus occurrence depended on the river's longitudinal profile, not on the chemical water quality, and that final success of introduction should be evaluated after several years.