The fellowship of the RING: the RING-B-box linker region interacts with the RING in TRIM21/Ro52, contains a native autoantigenic epitope in Sjögren syndrome, and is an integral and conserved region in TRIM proteins

Ro52 is a major autoantigen that is targeted in the autoimmune disease Sjögren syndrome and belongs to the tripartite motif (TRIM) protein family. Disease-related antigenic epitopes are mainly found in the coiled-coil domain of Ro52, but one such epitope is located in the Zn^2 +-binding region, which comprises an N-terminal RING followed by a B-box, separated by a ∼ 40-residue linker peptide. In the present study, we extend the structural, biophysical, and immunological knowledge of this RING-B-box linker (RBL) by employing an array of methods. Our bioinformatic investigations show that the RBL sequence motif is unique to TRIM proteins and can be classified into three distinct subtypes. The RBL regions of all three subtypes are as conserved as their known flanking domains, and all are predicted to comprise an amphipathic helix. This helix formation is confirmed by circular dichroism spectroscopy and is dependent on the presence of the RING. Immunological studies show that the RBL is part of a conformation-dependent epitope, and its antigenicity is likewise dependent on a structured RING domain. Recombinant Ro52 RING-RBL exists as a monomer in vitro, and binding of two Zn^2 + increases its stability. Regions stabilized by Zn^2 + binding are identified by limited proteolysis and matrix-assisted laser desorption/ionization mass spectrometry. Furthermore, the residues of the RING and linker that interact with each other are identified by analysis of protection patterns, which, together with bioinformatic and biophysical data, enabled us to propose a structural model of the RING-RBL based on modeling and docking experiments. Sequence similarities and evolutionary sequence patterns suggest that the results obtained from Ro52 are extendable to the entire TRIM protein family.     

Both acute and subchronic treatments with pindolol,a 5-HT1A and β1 and β2 adrenoceptor antagonist,elevate regional serotonin synthesis in the rat brain: An autoradiographic study

Antidepressant treatments, including those that increase serotonin (5-HT) neurotransmission, require several weeks or months until the onset of the therapeutic effect in depressed patients. The negative feedback on 5-HT transmission exhibited by the 5-HT_1A and 5-HT_1B autoreceptors has been postulated as a possible delaying factor. The aim of the present study was to assess the effect of the acute and subchronic treatment with pindolol, a 5-HT_1A/1B, β₁ and β₂ adrenoceptor antagonist, on 5-HT synthesis, one of the key parameters of 5-HT neurotransmission. Male Sprague–Dawley (SPD) rats (180–220 g) were treated with pindolol or an adequate volume of saline, administered either acutely (15 mg/kg i.p.; SPD-AC-SAL, SPD-AC-TR) or subchronically (15 mg/kg day i.p. for 7 days; SPD-SUBCHR-SAL, SPD-SUBCHR-TR). Thirty minutes following the single i.p. injection (acute experiment) or at the 8th day following the commencement of the subchronic treatment (subchronic experiment), 5-HT synthesis was measured using α-[¹⁴C]methyl-l-tryptophan autoradiography. The analysis of variance (ANOVA), followed by the Benjamini–Hochberg correction for multiple comparisons, revealed: (1) a significant increase of 5-HT synthesis in the SPD-AC-TR rats, relative to the SPD-AC-SAL rats in all brain regions examined except the substantia nigra – pars reticularis, dorsal subiculum, inferior olive, raphe magnus and raphe obscurus and (2) a significant increase of 5-HT synthesis in the SPD-SUBCHR-TR rats, relative to the SPD-SUBCHR-SAL rats in all brain regions except the median raphe, hypothalamus and raphe pontine. On the basis of these results, we hypothesized that the antagonism of the 5-HT_1A/1B receptors prevents the negative feedback mediated by these receptors on 5-HT synthesis, resulting in a persistent increase of 5-HT synthesis. The results accord with clinical reports on the utility of pindolol in the augmentation of antidepressant treatment.     

An acquisition trade-off with fast growth in a fish,the medaka <Emphasis Type=Italic>Oryzias latipes</Emphasis>: why do low-latitude ectotherms grow more slowly?

In various ectotherms, it is well-known that high-latitude individuals grow faster at any temperature than do low-latitude individuals as an adaptation to shorter growing seasons. The submaximal growth of low-latitude ectotherms implies that there exist trade-offs with fast growth. Although there is accumulating evidence for latitudinal variation in growth capacity, however, few studies have demonstrated these trade-offs. In this study, using a freshwater fish Oryzias latipes as a model organism, we tested whether there is an acquisition trade-off in which the higher growth capacity of high-latitude individuals is realized by foraging more intensively, despite the risk of predation. Foraging experiments revealed that higher-latitude O. latipes juveniles captured more prey per unit time than lower-latitude juveniles at any temperature. In addition, predation experiments revealed that higher-latitude juveniles were more frequently killed by dragonfly larvae. Observations of swimming behaviors demonstrated that the higher vulnerability to predation in high-latitude O. latipes is probably caused by lower steady- and burst-swimming performances due to the larger meal size, and partially by reduced caution due to a greater willingness to forage. We conclude that acquisition trade-offs may be the primary constraint on growth evolution across latitudes among fishes. This implies that growth capacity evolves in response not only to the length of the growing season but also to predation pressures.     

Morphological,molecular and phylogenetic characterisation of Eimeria macyi Wheat, 1975 (Apicomplexa: Eimeriidae) in the eastern red bat Lasiurus borealis (Müller) from Mississippi,USA

Systematic Parasitology - In November 2017, oöcysts of the coccidian Eimeria macyi Wheat, 1975 were isolated from the faeces of a single eastern red bat Lasiurus borealis Müller in...     

Mössbauer spectroscopy of the iron cores in human liver ferritin, ferritin in normal human spleen and ferritin in spleen from patient with primary myelofibrosis: preliminary results of comparative analysis

Comparative study of human liver ferritin and spleen tissues from healthy human and patient with primary myelofibrosis was carried out using Mössbauer spectroscopy with a high velocity resolution at 295 and 90 K and with a low velocity resolution at 20 K. The results obtained demonstrated that the iron content in patient’s spleen in the form of iron storage proteins was about ten times larger than that in normal tissue. However, in the case of patient with primary myelofibrosis the magnetic anisotropy energy barrier differed from that in normal case and, probably, the iron core size was supposed to be slightly larger than that in both normal spleen tissue and normal human liver ferritin in contrast to well-known data for iron overload in patients with thalassemia accompanied by the iron-core size increase. Therefore, the iron overload in the case of patient with primary myelofibrosis may be related to increase in the ferritin content mainly. It was also found that Mössbauer hyperfine parameters for normal and patient’s spleen and normal human liver ferritin demonstrated some small differences related, probably, to some small structural variations in the ferritin iron cores of patient’s spleen.     

Histological and molecular studies of species of Myxobolus Bütschli, 1882 (Myxozoa: Myxosporea) in the gills of Abramis, Blicca and Vimba spp. (Cyprinidae), with the redescription of M. macrocapsularis Reuss, 1906 and M. bliccae Donec & Tozyyakova, 1984

Although Myxobolus spp. from cyprinid fishes are generally characterised by a strict host-specificity, this study has found that the breams Abramis brama (L.), Blicca bjoerkna (L.) and Vimba vimba (L.) may be infected by the same Myxobolus spp. It is demonstrated that M. macrocapsularis Reuss, 1906, a parasite of the gill filaments of B. bjoerkna, can also infect A. brama. In the same way, M. bliccae Donec & Tozyyakova, 1984, also a parasite of B. bjoerkna, can also occur in V. vimba. The molecular sequences of M. macrocapsularis spores from B. bjoerkna and A. brama were 100% identical. Two of the 18S rDNA sequences of three replicate samples of M. bliccae from B. bjoerkna were 100% identical, whereas the third sequence exhibited a 99.7% similarity with sequences from V. vimba. M. bliccae sequences of spores collected from V. vimba showed a 99.8% similarity to the first two isolates and 99.6% to the third. Data obtained by morphological, histological and molecular biological methods all suggest that Myxobolus spp., known for their strict host-specificity, may sometimes infect several closely related cyprinids.     

The Comparison of the Outcomes between Primary PCI,Fibrinolysis, and No Reperfusion in Patients ≥ 75 Years Old with ST-Segment Elevation Myocardial Infarction: Results from the Chinese Acute Myocardial Infarction (CAMI) Registry

BackgroundOnly a few randomized trials have analyzed the clinical outcomes of elderly ST-segment elevation myocardial infarction (STEMI) patients (≥ 75 years old). Therefore, the best reperfusion strategy has not been well established. An observational study focused on clinical outcomes was performed in this population.MethodsBased on the national registry on STEMI patients, the in-hospital outcomes of elderly patients with different reperfusion strategies were compared. The primary endpoint was defined as death. Secondary endpoints included recurrent myocardial infarction, ischemia driven revascularization, myocardial infarction related complications, and major bleeding. Multivariable regression analysis was performed to adjust for the baseline disparities between the groups.ResultsPatients who had primary percutaneous coronary intervention (PCI) or fibrinolysis were relatively younger. They came to hospital earlier, and had lower risk of death compared with patients who had no reperfusion. The guideline recommended medications were more frequently used in patients with primary PCI during the hospitalization and at discharge. The rates of death were 7.7%, 15.0%, and 19.9% respectively, with primary PCI, fibrinolysis, and no reperfusion (P < 0.001). Patients having primary PCI also had lower rates of heart failure, mechanical complications, and cardiac arrest compared with fibrinolysis and no reperfusion (P < 0.05). The rates of hemorrhage stroke (0.3%, 0.6%, and 0.1%) and other major bleeding (3.0%, 5.0%, and 3.1%) were similar in the primary PCI, fibrinolysis, and no reperfusion group (P > 0.05). In the multivariable regression analysis, primary PCI outweighs no reperfusion in predicting the in-hospital death in patients ≥ 75 years old. However, fibrinolysis does not.ConclusionsEarly reperfusion, especially primary PCI was safe and effective with absolute reduction of mortality compared with no reperfusion. However, certain randomized trials were encouraged to support the conclusion.     

Both acute and subchronic treatments with pindolol,a 5-HT1A and β1 and β2 adrenoceptor antagonist,elevate regional serotonin synthesis in the rat brain: An autoradiographic study

Antidepressant treatments, including those that increase serotonin (5-HT) neurotransmission, require several weeks or months until the onset of the therapeutic effect in depressed patients. The negative feedback on 5-HT transmission exhibited by the 5-HT_1A and 5-HT_1B autoreceptors has been postulated as a possible delaying factor. The aim of the present study was to assess the effect of the acute and subchronic treatment with pindolol, a 5-HT_1A/1B, β₁ and β₂ adrenoceptor antagonist, on 5-HT synthesis, one of the key parameters of 5-HT neurotransmission. Male Sprague–Dawley (SPD) rats (180–220 g) were treated with pindolol or an adequate volume of saline, administered either acutely (15 mg/kg i.p.; SPD-AC-SAL, SPD-AC-TR) or subchronically (15 mg/kg day i.p. for 7 days; SPD-SUBCHR-SAL, SPD-SUBCHR-TR). Thirty minutes following the single i.p. injection (acute experiment) or at the 8th day following the commencement of the subchronic treatment (subchronic experiment), 5-HT synthesis was measured using α-[¹⁴C]methyl-l-tryptophan autoradiography. The analysis of variance (ANOVA), followed by the Benjamini–Hochberg correction for multiple comparisons, revealed: (1) a significant increase of 5-HT synthesis in the SPD-AC-TR rats, relative to the SPD-AC-SAL rats in all brain regions examined except the substantia nigra – pars reticularis, dorsal subiculum, inferior olive, raphe magnus and raphe obscurus and (2) a significant increase of 5-HT synthesis in the SPD-SUBCHR-TR rats, relative to the SPD-SUBCHR-SAL rats in all brain regions except the median raphe, hypothalamus and raphe pontine. On the basis of these results, we hypothesized that the antagonism of the 5-HT_1A/1B receptors prevents the negative feedback mediated by these receptors on 5-HT synthesis, resulting in a persistent increase of 5-HT synthesis. The results accord with clinical reports on the utility of pindolol in the augmentation of antidepressant treatment.     

The lectin from the mushroom Pleurotus ostreatus: a phosphatase-activating protein that is closely associated with an α-galactosidase activity: A part of this paper has been presented as a preliminary report at the 17th Interlec. Meeting 1997 in Würzburg,Germany

The lectin from the mushroom Pleurotus ostreatus described earlier [F. Conrad, H. Rüdiger, The lectin from Pleurotus ostreatus: purification, characterization and interaction with a phosphatase, Phytochemistry 36 (1994) 277–283] was further characterized. Determination of the isoelectric point by capillary electrophoresis gave a value of 7.6. The dissociation constant of the lectin-α-lactose-1-phosphate complex determined by capillary electrophoresis is 3 mM. The activation of an endogenous phosphatase by the lectin as found earlier for the pseudosubstrate p-nitrophenylphosphate was confirmed also for naturally occurring substrates as ADP and ATP. We observed that at all purification steps the lectin is accompanied by an α-galactosidase activity. Both activities could neither be resolved by electrophoresis under non-denaturing conditions nor by affinity chromatography. However, carbohydrate binding by the lectin and carbohydrate processing by the enzyme are not due to the same site since: (i) the lectin accepts both α- and β-glycosides whereas the enzyme activity is restricted to the α-anomer; (ii) the interaction with erythrocytes leads to a stable agglutinate, i.e. no ‘clot-dissolving activity’ [C.N. Hankins, J.I. Kindinger, L.M. Shannon, Legume α-galactosidases which have hemagglutinin properties, Plant Physiol. 65 (1980) 618–622] is observed; (iii) the α-galactosidase activity is inhibited by galactose but not by a β-galactoside. Therefore, lectin and enzymatic activities are either properties of two tightly associated proteins, or of just one molecule. The kinetic parameters of the lectin-associated α-galactosidase activity for p-nitrophenyl-α-galactopyranoside are: K_M=2.5 mM, k_cat=66 s⁻¹, and K_I=20 mM for the inhibitor d-galactose.     

Redescription of <Emphasis Type="Italic">Parspina argentinensis</Emphasis> (Szidat, 1954) (Digenea: Cryptogonimidae) from freshwater fishes (Pimelodidae) in the basins of the Paraná and La Plata Rivers,Argentina, with comments on <Emphasis Type="Italic">P. bagre</Emphasis> Pearse, 1920

The type-specimens of Parspina argentinensis (Szidat, 1954) from Pimelodus maculatus Lacépède, and new material from Iheringichthys labrosus (Lütken), Parapimelodus valenciennis (Lütken), Pimelodella gracilis (Valenciennes), Pimelodus albicans (Valenciennes), P. argenteus Perugia and P. maculatus caught in the basins of the La Plata and Paraná Rivers, Argentina, were studied to elucidate its taxonomic status. The type-specimens of Parspina bagre Pearse, 1920 from Pimelodella metae Eigenmann in Lake Valencia, Venezuela, were also studied and new observations on its morphology recorded. The amendation of the generic diagnosis of Parspina Pearse, 1920 is proposed based on the study of both species, which revealed, among other features, the absence of a thin walled-membrane enclosing the male terminal genitalia. Parspina argentinensis is characterised by the following features: absence of a gonotyl; presence of a bipartite seminal vesicle, pars prostatica and ejaculatory duct; caeca of nearly equal length; uterus extending from the level of the ventral sucker to end of body; testes symmetrical to slightly oblique; ovary transversely elongate, compact and variable in shape; and the utilisation of a wide range of freshwater fishes as hosts. The tegumental surface of this species is covered with pectinate spines arranged quincuncially. Spines decrease in size and density from forebody to hindbody. There are two types of sensory papillae, ciliated and dome-shaped. Ciliated papillae are distributed on the surface of the oral region and on the dorsal and ventral surfaces of the body, whereas dome-shaped papillae are found on the surface of the ventral rim of the oral sucker associated with a ciliated papilla. Gland-duct openings are interspersed with the spines of the oral crown. In addition, the infection indices of P. argentinensis vary widely, not only among different hosts but also between the Paraná and La Plata River basins. Higher prevalences of P. argentinensis are recorded in the La Plata River, and Pimelodus albicans, P. maculatus and Iheringichthys labrosus are the principal final hosts in this area.     

A new genus and species of turtle blood fluke (Digenea: Schistosomatoidea) from the Mekong snail-eating turtle, <Emphasis Type="Italic">Malayemys subtrijuga</Emphasis> (Schlegel &amp; Müller) (Testudines: Geoemydidae) in Vietnam,with a reassessment of related Asiatic turtle blood flukes and molecular phylogeny

Platt sinuosus Roberts & Bullard n. g., n. sp. (type-species) infects the kidney and mesenteric blood vessels of Mekong snail-eating turtles, Malayemys subtrijuga (Schlegel & Müller), in the Mekong River Basin. Species of Platt Roberts & Bullard n. g. are unique by the combination of having a papillate ventral sucker, vasa efferentia that are dorsal to the gonads, a massive cirrus-sac that is directed anteriad or laterad, and a vitellarium that surrounds the intestinal caeca. The new species resembles Platt ocadiae (Takeuti, 1942) Roberts & Bullard n. comb. but differs from it by having an external seminal vesicle that overlaps with or is immediately posterior to the level of the ventral sucker. Seven species previously of Hapalorhynchus Stunkard, 1922 are reassigned herein to Platt: P. odhnerensis (Mehra, 1933) Roberts & Bullard n. comb.; P. yoshidai (Ozaki, 1939) Roberts & Bullard n. comb.; P. ocadiae; P. oschmarini (Belous, 1963) Roberts & Bullard n. comb.; P. sutlejensis (Mehrotra, 1973) Roberts & Bullard n. comb.; P. synderi (Platt & Sharma, 2012) Roberts & Bullard n. comb.; and P. tkachi (Platt & Sharma, 2012) Roberts & Bullard n. comb. A dichotomous key to Platt spp. is provided. Hapalorhynchus sheilae (Mehrotra, 1973) Bourgat, 1990 and Hapalorhynchus mica (Oshmarin, 1971) Bourgat, 1990 are considered as species inquirendae, and Hapalorhynchus indicus (Thapar, 1933) Price, 1934 and Hapalorhynchus macrotesticularis (Rohde, Lee, & Lim, 1968) Brooks & Sullivan, 1981 are considered as species incertae sedis. Phylogenetic analysis of the large subunit rDNA (28S) showed P. sinuosus and P. snyderi to be sister taxa distinct from a monophyletic Hapalorhynchus and Coeuritrema platti Roberts & Bullard, 2016.