Structure of the haptoglobin-haemoglobin complex

Red cell haemoglobin is the fundamental oxygen-transporting molecule in blood, but also a potentially tissue-damaging compound owing to its highly reactive haem groups. During intravascular haemolysis, such as in malaria and haemoglobinopathies(1), haemoglobin is released into the plasma, where it is captured by the protective acute-phase protein haptoglobin. This leads to formation of the haptoglobin-haemoglobin complex, which represents a virtually irreversible non-covalent protein-protein interaction(2). Here we present the crystal structure of the dimeric porcine haptoglobin-haemoglobin complex determined at 2.9 angstrom resolution. This structure reveals that haptoglobin molecules dimerize through an unexpected beta-strand swap between two complement control protein (CCP) domains, defining a new fusion CCP domain structure. The haptoglobin serine protease domain forms extensive interactions with both the alpha- and beta-subunits of haemoglobin, explaining the tight binding between haptoglobin and haemoglobin. The haemoglobin-interacting region in the alpha beta dimer is highly overlapping with the interface between the two alpha beta dimers that constitute the native haemoglobin tetramer. Several haemoglobin residues prone to oxidative modification after exposure to haem-induced reactive oxygen species are buried in the haptoglobin-haemoglobin interface, thus showing a direct protective role of haptoglobin. The haptoglobin loop previously shown to be essential for binding of haptoglobin-haemoglobin to the macrophage scavenger receptor CD163 (ref. 3) protrudes from the surface of the distal end of the complex, adjacent to the associated haemoglobin alpha-subunit. Small-angle X-ray scattering measurements of human haptoglobin-haemoglobin bound to the ligand-binding fragment of CD163 confirm receptor binding in this area, and show that the rigid dimeric complex can bind two receptors. Such receptor cross-linkage may facilitate scavenging and explain the increased functional affinity of multimeric haptoglobin-haemoglobin for CD163 (ref. 4).

Microbial transformation of the sesquiterpene lactone tagitinin C by the fungus Aspergillus terreus

The biotransformation of the sesquiterpene lactone tagitinin C by the fungus Aspergillus terreus MT 5.3 yielded a rare derivative that was elucidated by spectrometric methods. The fungus led to the formation of a different product through an unusual epoxidation reaction between C4 and C5, formation of a C3,C10 ether bridge, and a methoxylation of the C1 of tagitinin C. The chemical structure of the product, namely 1 beta-methoxy-3 alpha-hydroxy-3,10 beta-4,5 alpha-diepoxy-8 beta-isobutyroyloxygermacr-11(13)-en-6 alpha,12-olide, is the same as that of a derivative that was recently isolated from the flowers of a Brazilian population of Mexican sunflower (Tithonia diversifolia), which is the source of the substrate tagitinin C. The in vitro cytotoxic activity of the substrate and the biotransformed product were evaluated in HL-60 cells using an MTT assay, and both compounds were found to be cytotoxic. We show that soil fungi may be useful in the biotransformation of sesquiterpene lactones, thereby leading to unusual changes in their chemical structures that may preserve or alter their biological activities, and may also mimic plant biosynthetic pathways for production of secondary metabolites.

Sympathoexcitation during chemoreflex active expiration is mediated by L-glutamate in the RVLM/Botzinger complex of rats

Moraes DJ, Zoccal DB, Machado BH. Sympathoexcitation during chemoreflex active expiration is mediated by L-glutamate in the RVLM/Botzinger complex of rats. J Neurophysiol 108: 610-623, 2012. First published April 25, 2012; doi:10.1152/jn.00057.2012.-The involvement of glutamatergic neurotransmission in the rostral ventrolateral medulla/Botzinger/pre-Botzinger complexes (RVLM/BotC/pre-BotC) on the respiratory modulation of sympathoexcitatory response to peripheral chemoreflex activation (chemoreflex) was evaluated in the working heart-brain stem preparation of juvenile rats. We identified different types of baro- and chemosensitive presympathetic and respiratory neurons intermingled within the RVLM/BotC/pre-BotC. Bilateral microinjections of kynurenic acid (KYN) into the rostral aspect of RVLM (RVLM/BotC) produced an additional increase in frequency of the phrenic nerve (PN: 0.38 +/- 0.02 vs. 1 +/- 0.08 Hz; P < 0.05; n = 18) and hypoglossal (HN) inspiratory response (41 +/- 2 vs. 82 +/- 2%; P < 0.05; n = 8), but decreased postinspiratory (35 +/- 3 vs. 12 +/- 2%; P < 0.05) and late-expiratory (24 +/- 4 vs. 2 +/- 1%; P < 0.05; n = 5) abdominal (AbN) responses to chemoreflex. Likewise, expiratory vagal (cVN; 67 +/- 6 vs. 40 +/- 2%; P < 0.05; n = 5) and expiratory component of sympathoexcitatory (77 +/- 8 vs. 26 +/- 5%; P < 0.05; n = 18) responses to chemoreflex were reduced after KYN microinjections into RVLM/BotC. KYN microinjected into the caudal aspect of the RVLM (RVLM/pre-BotC; n = 16) abolished inspiratory responses [PN (n = 16) and HN (n = 6)], and no changes in magnitude of sympathoexcitatory (n = 16) and expiratory (AbN and cVN; n = 10) responses to chemoreflex, producing similar and phase-locked vagal, abdominal, and sympathetic responses. We conclude that in relation to chemoreflex activation 1) ionotropic glutamate receptors in RVLM/BotC and RVLM/pre-BtC are pivotal to expiratory and inspiratory responses, respectively; and 2) activation of ionotropic glutamate receptors in RVLM/BotC is essential to the coupling of active expiration and sympathoexcitatory response.

Compression stockings have a synergistic effect with walking in the late afternoon to reduce edema of the lower limbs

Aim. The aim of this study was to evaluate whether an association of elastic stockings and walking for a short period in the late afternoon reduces leg edema. Methods. Volume changes of the legs of sixteen patients (32 limbs), who walked on a treadmill for 30 minutes using elastic compression stockings, were analyzed in a quantitative, cross-over randomized (in order of arrival at the clinic) study. They were submitted to volumetry using the water displacement technique and subsequently required to put on 20/30 made-to-measure compression stockings (Sigvaris). The patients walked on a treadmill for 30 minutes and after removing the stockings volumetry of the legs was again performed. Legs were assessed using the CEAP classification and divided into groups. Analysis of variance was used for statistical analysis with an alpha error of 5% being considered acceptable. Results. When participants walked wearing compression stockings, there was a reduction in leg volume. When the CEAP classification was evaluated, it was noted that there was a statistically significant difference for the CEAP C0, C1 and C2 categories of legs using stockings compared to those that did not use. Conclusion. Compression stockings have a synergistic effect with walking in the late afternoon thus reducing edema of the lower limbs. [Int Angiol 2012;31:490-3]

Resiliência física de dois latossolos vermelhos sob plantio direto

A resiliência física de solos é proveniente de processos regenerativos que incluem ciclos de umedecimento e secamento, congelamento e descongelamento, assim como as atividades biológicas. Este estudo testou a hipótese de que as propriedades físicas do solo, como a permeabilidade do solo ao ar, densidade do solo, porosidade de aeração e porosidade total, são indicadores físicos eficientes para quantificar a resiliência de solos de diferentes texturas submetidos ao estresse mecânico (compactação) e após subsequentes ciclos de umedecimento e secamento. O objetivo foi avaliar o comportamento e a resiliência do solo por meio de propriedades físicas de dois Latossolos Vermelhos. Foram retiradas 25 amostras indeformadas (0,00-0,05 m) de dois solos: solo I, com textura argilosa, e solo II, com textura franco-argilo-arenosa, realizando as determinações das propriedades físicas nos tratamentos: antes da compactação (A), depois da compactação (C0) e após ciclos de umedecimento e secamento (C1, C2, C3 e C4). As propriedades densidade do solo e porosidade total não apresentaram recuperação da condição inicial após a compactação nos solos I e II; as propriedades conteúdo volumétrico de água e porosidade de aeração mostraram recuperação parcial apenas no solo I; e a permeabilidade do solo ao ar foi a propriedade que apresentou a melhor recuperação e a maior resiliência. Em relação ao distinto comportamento dos dois solos, observou-se que o solo I foi mais resiliente que o solo II nas propriedades que apresentaram recuperação.

A uniqueness theorem for the determination of sources in the Germain–Lagrange plate equation

We prove a uniqueness result related to the Germain–Lagrange dynamic plate differential equation. We consider the equation {∂2u∂t2+△2u=g⊗f,in ]0,+∞)×R2,u(0)=0,∂u∂t(0)=0, where uu stands for the transverse displacement, ff is a distribution compactly supported in space, and g∈Lloc1([0,+∞)) is a function of time such that g(0)≠0g(0)≠0 and there is a T0>0T0>0 such that g∈C1[0,T0[g∈C1[0,T0[. We prove that the knowledge of uu over an arbitrary open set of the plate for any interval of time ]0,T[]0,T[, 0

Hypervitaminosis A-induced hepatic fibrosis in a cat

Rationale: The excessive intake of vitamin A in the form of vitamin concentrate, supplement or vitamin-rich liver can result in hypervitaminosis A in man and animals. Although osteopathologies resulting from chronic vitamin A intoxication in cats are well characterized, no information is available concerning feline hypervitaminosis A-induced liver disease. Clinical summary: We report the first case of hepatic stellate cell lipidosis and hepatic fibrosis in a domestic cat that had been fed a diet based on raw beef liver. Radiographic examination revealed exostoses and ankylosis between vertebrae C1 and T7, compatible with deforming cervical spondylosis. Necropsy showed a slightly enlarged and light yellow to bronze liver. Microscopic and ultrastructural analyses of liver tissues revealed diffuse and severe liver fibrosis associated with hepatic stellate cell hyperplasia and hypertrophy. These cells showed immunopositive staining for α-smooth muscle actin and desmin markers. The necropsy findings of chronic liver disease coupled with osteopathology supported the diagnosis of hypervitaminosis A. Practical relevance: As in human hepatology, if there is dietary evidence to support increased intake of vitamin A, then hypervitaminosis A should be considered in the differential diagnosis of chronic liver disease in cats.